Abstract:
The invention concerns a pharmaceutical composition for treating or preventing C hepatitis (HCV), induced infections, which in a preferred embodiment, comprises a main active principle, (i) a fusion polypeptide, including the HCV capsid polypeptide (C191) and polypeptide coat (E1) and in which at least one cleavage site 173/174 and 191/192 has been made inoperative by mutation; (ii) an equimolar mixture of the C191 polypeptide of which the cleavage site 173/174 has been made inoperative and of the E1 polypeptide (mixture equivalent to the fusion polypeptide); or (iii) a DNA molecule coding for this fusion polypeptide. Products (i) to (iii) are characterized in that the C191 element is incapable of regulating the functioning of the genes, in particular of causing them to interact. Such a composition can also include any form equivalent to the products described above.

Description:
RELATED APPLICATIONS  
       [0001]    This application is a divisional of application Ser. No. 09/388,874, filed Sep. 2, 1999, which is a continuation-in-part of International Application No. PCT/FR98/00448, which designated the United States and was filed on Mar. 6, 1998, published in French, which claims priority to a French Application 97/02,887, filed on Mar. 6, 1997. The entire teachings of the above applications are incorporated herein by reference. 
     
    
     
       BACKGROUND OF THE INVENTION  
         [0002]    The hepatitis C virus (HCV) is the agent responsible for the majority of hepatitis infections of the non-A non-B type. The seroprevalence of HCV infections varies between 0.3 abd 1.5% in the world population, possibly reaching 18% in some developing countries. Hundreds of millions of people are thus thought to be infected worldwide. Nine types and thirty subtypes of HCV have been described. The subtypes may be associated with a defined geographical distribution, type 1b being the most widespread worldwide. The progression to the chronic form occurs in 50% of cases, about 5 years after the primary infection. Persistent Chronic Hepatitis which is asymptomatic, but which exhibits a high circulating virus titer, is first observed, then Active Chronic Hepatitis becomes established. Twenty percent of chronic hepatitis progress to sclerosis of the liver within about ten years. Hepatocarcinoma may develop in the cirrhotic liver.  
         SUMMARY OF THE INVENTION  
         [0003]    The present invention relates to a nucleic acid molecule that encodes a fusion polypeptide that comprises a first region consisting essentially of the C polypeptide of the hepatitis C virus (HCV) or a portion thereof that comprises a polypeptide region responsible for gene regulatory activity; and a second region consisting essentially of the envelope polypeptide (E1) of the virus or a portion thereof that comprises a site for cytoplasmic anchorage of the E1 polypeptide. The first region is fused by a peptide bond to the second region, and the fusion polypeptide is not cleaved by a mammalian protease.  
           [0004]    In one embodiment, the present invention also pertains to a nucleic acid molecule that encodes a fusion polypeptide that has a C polypeptide of HCV or a portion thereof that comprises a first C polypeptide region responsible for gene regulatory activity and a second C polypeptide region responsible for the interaction with the E1 envelope polypeptide, wherein the site of interaction with the E1 polypeptide is between about 151 and about 173, or between about 173 and about 191. The fusion polypeptide also contains a E1 envelope polypeptide of HCV or a portion thereof that comprises a first E1 polypeptide region responsible for E1 cytoplasmic anchorage, and a second E1 polypeptide region responsible for the interaction of said second C polypeptide region, wherein the site for interaction with the C polypeptide is between about amino acid 330 and about amino acid 380. The C polypeptide is fused by a peptide bond to said E1 envelope polypeptide, and the C polypeptide comprises Cysteine 172 -Serine 173 -Phenylalanine 174 -Serine 175  with at least one mutation between amino acid Nos 172 and 175. In a particular embodiment, the Serine residue in position 173 can be substituted by a Methionine residue or the Phenylalanine residue can be substituted by a Leucine residue.  
           [0005]    The present invention also embodies a mixture that comprises a first polypeptide comprising at least a portion of the C polypeptide of HCV having a site for interaction with the E1 polypeptide of HCV; and a second polypeptide comprising at least a portion of the E1 polypeptide of HCV having a site for interaction with the C polypeptide of HCV and a site for cytoplasmic anchorage of the E1 polypeptide. In another embodiment the present invention includes a mixture that comprises a first polypeptide consisting essentially of the C polypeptide of the hepatitis C virus (HCV) or a portion thereof that comprises a polypeptide region responsible for gene regulatory activity; and a second polypeptide consisting essentially of the envelope polypeptide (E1) of the virus or a portion thereof that comprises a site for cytoplasmic anchorage of the E1 polypeptide. The mixture is not cleaved by a mammalian protease. The first and second polypeptides can be in a quantity that is substantially equimolar.  
           [0006]    The present invention includes vaccine and/or pharmaceutical compositions comprising the nucleic acid sequences, polypeptide sequences or mixtures described herein. The present invention also relates to plasmids, vectors and cells that comprise the nucleic acid sequence of the present invention, or that encode the polypeptide sequence of the present invention.  
           [0007]    The present invention also embodies methods of treating an individual having HCV, methods for preventing and individual from contracting or becoming affected with HCV, methods of vaccinating an individual against HCV, and methods for eliciting an immune response in an individual, comprising administering to the individual the nucleic acid molecules or polypeptide molecules described herein. 
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0008]    [0008]FIG. 1 is a schematic representation of the HCV genome which consists of RNA with its untranslated 5′ and 3′ regions indicated by lines, and the open reading frame of the precursor polyprotein indicated in the form of a rectangle.  
         [0009]    [0009]FIG. 2 represents the inserts derived from the HCV genome which are tested in plasmids pRC. The sequences derived from the HCV genome are represented by a rectangle and the mutated residues are indicated by dots.  
         [0010]    [0010]FIG. 3 is a diagram representing the luciferase activity measured for each of the constructs of which some are mutated at the level of one or more cleavage sites. The identity of the insert tested appears on the x-axis while the quantity of luciferase produced relative to the total quantity of protein produced appears on the y-axis.  
         [0011]    FIGS.  4 A- 4 L are an illustration of the nucleic acid sequence (SEQ ID NO.: 1) and amino acid sequence (SEQ ID NO.: 2) of HCV. 
     
    
     DETAILED DESCRIPTION OF THE INVENTION  
       [0012]    The invention relates to a pharmaceutical composition intended for the treatment or prophylaxis of infections induced by the hepatitis C virus (HCV).  
         [0013]    The hepatitis C virus (HCV) is a positive single-stranded RNA virus. On the basis of structural resemblance, HCV has been linked to the flavivirus and pestivirus families.  
         [0014]    During an infectious event, the HCV genome is first translated into a precursor polyprotein of about 3000 amino acids. This polyprotein then undergoes post-translational cleavages to give various precursors and mature viral proteins. The structural proteins of HCV are located in the N-terminal region of the polyprotein. As shown in FIG. 1, they are more particularly the capsid or core protein (C), and the envelope proteins E1 and E2, which are present in the following order: NH2-C-E1-E2. This portion is cleaved by the host cell proteases.  
         [0015]    The numbering of the amino acids of the polyprotein as well as of its derivatives, which is adopted hereafter, is that commonly used and in particular presented by Choo et al., PNAS [vol. 88: p. 2451 (1991)]. Thus, the C protein corresponds to the amino acids at positions 1 to 191 of the polyprotein and the E1 protein to the amino acids at positions 192 to 380. In the remainder of the text, it is appropriate to number the amino acids of the sequence of the E1 protein, from position 192 to position 380, 381, 382 or 383. In the remainder of the text, for the sake of simplicity, reference is made solely to the C-terminal position 380.  
         [0016]    The C protein derived from the direct cleavage of the polyprotein contains 191 amino acids. This C protein, also called C191, may itself be truncated toward its C-terminal end by enzymatic cleavage to give a protein of 173 amino acids, called C173. In the remainder of the test, the term “C protein or polypeptide” will preferably designate the C191 form.  
         [0017]    The C protein is a good vaccine candidate since it is of course a structural protein of the virus and since the region encoding this protein is relatively well conserved by the various HCV strains. It is known that a region of the C protein capable of generating a high antibody response corresponds to the first 120 amino acids; the first 48 amino acids constituting the major antigenic domain. However, a major obstacle to its use as vaccine lies in the fact that this protein is capable of transactivating genes belonging to the host cell, in particular genes such a oncogenes, which may have, inter alia, the consequence of inducing a carcinogenesis event.  
         [0018]    Indeed, it has in particular been shown that the C173 form was capable of translocation in the nucleus of the host cell and of transactivation. The region of the C protein responsible for the translocation in the nucleus and for the regulatory acitvity appears to be located in the N-terminal portion (first 123 amino acids).  
         [0019]    Thus, the region of the C protein which is of interest from a vaccine point of view is, on the other hand, responsible for a toxic effect toward the host cell.  
         [0020]    To overcome this difficulty, a solution commonly envisaged in the scientific community would be to use a C191 protein whose cleavage site at position 173/174 would have been made inoperative by mutation. As will be seen below, such a protein nevertheless proves capable of regulatory activity, even if it is to a lesser degree.  
         [0021]    Surprisingly, it has now been demonstrated that it was possible to abolish the regulatory activity of the C protein by modifying it and by combining it, under certain conditions, with the E1 protein. The present invention provides means for abolishing the regulatory activity by preventing the migration of the C protein into the nucleus. This migration no longer takes place in the presence of the E1 protein which possesses, inter alia, the property of becoming anchored in the cytoplasm, at the level of the endoplasmic reticulum, and which, unexpectedly, has the capacity to retain the C protein therein when certain conditions are met. The migration may be abolished by producing, for example, a fusion of the two proteins, cleavable or otherwise; in the case where it is cleavable, the products generated should be capable of interacting with each other so that there is no leakage of one of them into the nucleus; the complex formed by the product of cleavage being capable of becoming anchored in the cytoplasm. The equivalent of a cleavable peptide fusion is a mixture, in equimolar quantity, of the components constituting the fusion.  
         [0022]    Accordingly, the subject of the invention is a pharmaceutical composition comprising:  
         [0023]    (i) A polypeptide which contains:  
         [0024]    (a) a first region corresponding to all or part of the C polypeptide of the hepatitis C virus; and  
         [0025]    (b) a second region corresponding to all or part of the E1 polypeptide of said virus and, proves, as such or via its products of cleavage, incapable of regulatory activity toward one or more genes;  
         [0026]    (ii) A mixture (preferably) in substantially equimolar quantity,  
         [0027]    (a) of a first polypeptide containing a region which corresponds to all or part of the C polypeptide of HCV and  
         [0028]    (b) of a second polypeptide containing a region corresponding to all or part of the E1 polypeptide of HCV; and  
         [0029]    which proves incapable of regulatory activity toward one or more genes; or  
         [0030]    (iii) A DNA molecule comprising a sequence encoding the polypeptide as described in (i) of the present claim, placed under the control of elements necessary for its expression in a mammalian cell; and a pharmaceutically acceptable carrier or diluent.  
         [0031]    (iv) According to another aspect of the invention, the subject of the invention is also a method for the treatment or prevention of an infection induced by HCV according to which a pharmaceutical composition according to the invention is administered to a mammal, preferably a human, requiring such a treatment.  
         [0032]    (v) “Polypeptide” is understood to mean any chain of amino acids covalently linked to each other, regardless of the length of the chain and regardless of the post-translation modifications which may take place such as, for example, a lipidation. It is also possible to use the term protein interchangeable.  
         [0033]    (vi) “C polypeptide of HCV” is understood to mean in particular a C polypeptide which possesses the amino acid sequence as disclosed by Choo, et al. as well as any other C polypeptide obtained from any other strain and whose sequence could differ from that of Choo, et al. For example, it may represent the C polypeptides described by Takeuchi, K., et al., (Nucleic Acids Research 18: 4626 (1990)); Houghton, M., et al. (Hepatology 14:381 (1991)); Delisse, et al. (J. Hepatoloty, 13, suppl. 4 (1991)); Bukh, J., et al., (PNAS 91:8239 (1994)); and Hiroaki, O., et al., (Intervirology 37:68 (1994)).  
         [0034]    “E1 polypeptide of HCV” is understood to mean in particular an E1 polypeptide which possesses the amino acid sequence as disclosed by Choo, et al., as well as any other E1 polypeptide obtained from any other strain and whose sequence could differ from that of Choo, et al. For example, it may represent the E1 polypeptides described in Hiroaki, O., et al. (Intervirology 37: 68 (1994)), Grakoui, et al., (J. Virol. 67:1385 (1993)); and Spaete, et al. (Virology 188:819 (1992)); Matsumia, et al. (J. Virol. 66: 1425); or in Kohara, et al. (J. Gen. Virol. 73:2313 (1992)).  
         [0035]    Thus, the amino acid sequence of the C polypeptide and that of the E1 polypeptide of HCV may vary according to the viral strain, reflecting the phenomenon of allelic variance. For example, a virus is usually represented by a set of strains which differ from each other in minor allelic characteristics. A polypeptide which fulfills the same biological function in different strains may have an amino acid sequence which is not the same for all the strains. Such an allelic variation is also found at the level of the DNA.  
         [0036]    At the level of the amino acid sequence, the allelic differences may consist of one or more amino acid substitutions, deletions or additions which do not alter the biological function.  
         [0037]    As regards the polypeptide included in the pharmaceutical composition according to the invention, two cases must be envisaged: either the polypeptide is incapable of being cleaved by a protease in a mammalian cell, or it is susceptible to such a cleavage.  
         [0038]    When the polypeptide is incapable of being cleaved by a protease in a mammalian cell, it advantageously contains:  
         [0039]    (a) a first region corresponding at least to the portion of the C polypeptide of the HCV virus responsible for the regulatory activity of said C polypeptide toward one or more genes; and  
         [0040]    (b) a second region corresponding at least to a portion of the E1 polypeptide of said virus responsible for the cytoplasmic anchorage of the E1 polypeptide.  
         [0041]    When the polypeptide is capable of being cleaved by a protease in a mammalian cell, it advantageously contains:  
         [0042]    (a) a first region corresponding at least to a portion of the C polypeptide of HCV responsible for the regulatory activity of said C polypeptide toward one or more genes and to the portion of said C polypeptide responsible for the interaction of said C polypeptide with the E1 polypeptide of said virus; and  
         [0043]    (b) a second region corresponding at least to a portion of the E1 polypeptide of said virus responsible for the interaction of the E1 polypeptide with the C polypeptide of said virus and to a portion of the E1 polypeptide of said virus responsible for the cytoplasmic anchorage of the E1 polypeptide.  
         [0044]    “Portion of the C polypeptide of HCV responsible for the regulatory activity of said C polypeptide toward one or more genes” is understood to mean in particular any portion of the C polypeptide of HCV capable of activating, transactivating or suppressing the transcription or the expression of any gene, according to any mechanism. This gene may be an eukaryotic gene, a viral gene, an oncogene or a protooncogene.  
         [0045]    A portion of the C polypeptide of HCV responsible for the regulatory activity of said polypeptide may in particular correspond to the amino acids at positions 38 to 43, 58 to 64, 66 to 71, 6 to 23, 39 to 74, 99 to 102, 101 to 121, 101 to 122, 58 to 121, 1 to 120, 1 to 121, 1 to 122, 1 to 123 or 1 to 173. Preferably, a portion of the C polypeptide of HCV responsible for thr regulatory activity may be a portion of the C polypeptide ranging from the amino acid at position 1 to the amino acid in one of positions 48 to 191. For this purpose, one of positions 48 to 191 may be for example position 119, 120, 121, 123 and 173.  
         [0046]    A portion of the C polypeptide of HCV responsible for the interaction of said C polypeptide with the E1 protein of HCV may in particular correspond to the amino acids at positions 151 to 173 or at positions 173 to 191 of the C polypeptide of HCV.  
         [0047]    A portion of the E1 polypeptide of HCV responsible for the cytoplasmic anchorage of the E1 polypeptide may be a hydrophobic domain of the E1 polypeptide. Such hydrophobic domains are for example located at positions 262 to 291, 370 to 380 and 330 to 380 of the E1 polypeptide.  
         [0048]    A portion of the E1 polypeptide of HCV responsible for the interaction of the C polypeptide with the E1 protein may be in particular the C-terminal domain of the E1 polypeptide, preferably the domain at positions 330 to 380 or at positions 370 to 380.  
         [0049]    In a polypeptide useful for the purposes of the present invention, the first region may be located on the N- or C-terminal side of the polypeptide, advantageously on the N-terminal side; likewise, the second region may be located on the C- or N-terminal side, advantageously on the C-terminal side. According to a preferred mode, the C-terminal end of the first region may be fused by peptide bonding to the N-terminal end of the second region.  
         [0050]    When the polypeptide contained in the pharmaceutical composition according to the invention comprises the region corresponding at least to the amino acids at positions 172 to 175 of the C polypeptide of HCV, this polypeptide advantageously contains a mutation making the cleavage site at position 173/174 inoperative. According to a preferred mode, such a mutation is a point mutation, carried out in one of positions 172 to 175. It may be obtained, for example, by deletion, addition or substitution of one or more amino acids, in particular by deletion, addition or substitution of one or more amino acid at positions 172 to 175. Preferably the mutation will be produced by substitution of one or two amino acids; a double mutation by substitution being most particularly preferred. According to a particular example, the residue naturally existing at positions 173 (serine) may be in particular substituted by the methionine residue and the residue naturally existing at position 173 (phenylalanine) may be substituted in particular by the leucine residue. h general, it is within the capability of persons skilled in the art to produce one or more mutations capable of making inoperative the cleavage site at position 173/174 of the C polypeptide of HCV.  
         [0051]    When the polypeptide contained in the pharmaceutical composition according to the invention comprises the region corresponding at least to the amino acids at positions 190 to 193 of the HCV polyprotein, this polypeptide advantageously contains a mutation making inoperative the cleavage site at position 191/192 of the HCV polyprotein. According to a preferred mode, such a mutation is a point mutation produced in one of positions 190 to 193. It may be obtained, for example, by deletion, addition or substitution of one or more amino acids, in particular by deletion, addition or substitution of one or more amino acids at positions 190 to 193. Preferably, the mutation will be produced by substitution of one or two amino acids, a double mutation by substitution being most particularly preferred. According to a specific example, the residue naturally existing at position 191 (alanine) may in particular be substituted by the valine residue and the residue naturally existing at position 192 (tyrosine) may in particular be substituted by the asparagine residue. In general, it is within the capabiity of persons skilled in the art to produce one or more mutations capable of making inoperative the cleavage site at position 191/192.  
         [0052]    When the polypeptide useful for the purposes of the present invention comprises both the region corresponding at least to amino acids 190 to 193 of the HCV polyprotein and region corresponding at least to amino acids 172 to 175 of the C polypeptide of HCV, only one of the two cleavage sites 191/192 and 173/174 can be made inoperative, preferably both will be made inoperative. When only the site 173/174 is made inoperative, the polypeptide is capable of being cleaved and in this particular case, it is necessary that this polypeptide possesses a first region which corresponds, inter alia, to the portion of the C polypeptide responsible for the interaction of said polypeptide with the E1 polypeptide and a second region which corresponds, inter alia, to the portion of the E1 polypeptide responsible for the interaction of said polypeptide with the C polypeptide.  
         [0053]    When a polypeptide useful for the purposes of the present invention is incapable of being cleaved by a protease, it may contain a cleavage site on the condition, however, that this cleavage site is not functional. For example, in the particular case of a polypeptide consisting of the C191 polypeptide fused with the E1 polypeptide and containing a mutation making inoperative the cleavage site at position 191/192, the cleavage site at position 173/174 may not be mutated; however, it will not be, or will be only slightly, functional, insofar as the cleavage at position 191/192 is no longer possible. Indeed, it is known that the cleavage at position 191/192 must be carried out for the cleavage at position 173/174 to take place.  
         [0054]    According to a specific mode, a polypeptide useful for the purposes of the present invention is incapable of being cleaved by a protease and contains:  
         [0055]    (a) a first region which substantially corresponds to the domain of the C polypeptide ranging from the amino acid at position 1 to the amino acid in one of positions 120 to 173, and  
         [0056]    (b) a second region which substantially corresponds to a domain of the E1 polypeptide containing at least one hydrophobic region, for example to the domain of the E1 polypeptide ranging from the amino acid at position 192 to the amino acid at position 380, or from the amino acid at position 330 to the amino acid at position 380, or from the amino acid at position 260 to the amino acid at position 290, or from the amino acid at position 260 to the amino acid at position 380.  
         [0057]    According to another particular mode, a polypeptide useful for the purposes of the present invention is incapable of being cleaved by a protease and contains:  
         [0058]    (a) a first region which substantially corresponds to the domain of the C polypeptide ranging from the amino acid at position 1 to the amino acid inone of positions 120 to 191, and  
         [0059]    (b) a second region which substantially corresponds to a domain of the E1 polypeptide containing at least one hydrophobic region, for example to the domain of the E1 polypeptide ranging from the amino acid at position 192 to the amino acid at position 380, or from the amino acid at position 330 to the amino acid at position 380, or from the amino acid at position 260 to the amino acid at position 290, or from the amino acid at position 260 to the amino acid at position 380;  
         [0060]    on the condition that said polypeptide does not contain a cleavage site 191/192 or alternatively when the cleavage site is reconstituted, then a mutation is introduced in order to make it inoperative.  
         [0061]    Advantageously, the first region of the polypeptide useful for the purposes of the present invention corresponds to the amino acids at positions 1 to 191 of the C polypeptide of HCV and/or the second region of this polypeptide corresponds at least to the amino acids at positions 192 to 380 of the E1 polypeptide of HCV. In a particularly preferred manner, the first and second regions are as defined above in this same paragraph, the amino acid at position 191 being fused by peptide bonding to the amino acid at position 192. According to a particular mode, the polypeptide consists of the first and second regions as defined above in this same paragraph.  
         [0062]    When the polypeptide useful for the purposes of the present invention is as described in the preceding paragraph, it imperatively contains a mutation makine inoperative the cleavage site at position 191/192 or at position 173/174. Preferably, the two cleavage sites are made inoperative.  
         [0063]    A mixture useful for the purposes of the present invention advantageously comprises:  
         [0064]    (a) a first polypeptide containing a region which corresponds at least to the portion of the C polypeptide of the HCV virus responsible for the regulatory activity of said C polypeptide toward one or more genes and to the portion of said C polypeptide responsible for the interaction of said C polypeptide with the E1 polypeptide of said virus, and  
         [0065]    (b) a second polypeptide containing a region corresponding to a portion of the E1 polypeptide of said virus responsible for the interaction of the E1 polypeptide with the C polypeptide of said virus and to a portion of the E1 polypeptide of said virus responsible for the cytoplasmic anchorage of the E1 polypeptide.  
         [0066]    The portions of the C and E1 polypeptides responsible for the properties listed in points (a) and (b) of the preceding paragraph may be as described above for the fusion polypeptide.  
         [0067]    Preferably, the first polypeptide of the mixture contains and in a most particularly preferred manner consists of a region corresponding to the amino acids at positions 1 to 191 of the C polypeptide (C191) of HCV. In the latter case, the cleavage site at position 173/174 must be made inoperative by mutation. This mutation may be produced as described above for the fusion polypeptide.  
         [0068]    Preferably, the second polypeptide of the mixture contains and in a most particularly preferred manner consists of a region corresponding to the amino acids at positions 192 to 380 of the E1 polypeptide of HCV.  
         [0069]    For the purposes of the present invention, a DNA molecule may be a simple linear DNA fragment, or alternatively a plasmid or alternatively a viral vector such as a pox vector.  
         [0070]    A polypeptide, a mixture or a molecule of DNA as described in the present application are of a most special interest when they are used for the manufacture of a medicament intended for the treatment or prevention of infections induced by HCV. They are in particular useful in the immunotherapy of infections induced by HCV, most particularly a DNA molecule.  
         [0071]    Finally, the invention relates to a method for inducing an immune response toward HCV in a mammal, according to which an immunologically effective quantity of a composition according to the invention is administered to said mammal in order to develop an immune response. The invention also relates to a method for the prevention or treatment of an infection induced by HCV, according to which a prophylactically or therapeutically effective quantity of a composition according to the invention is administered to an individual.  
         [0072]    The methods and the pharmaceutical compositions according to the invention can treat or prevent HCV infections and consequently hepatic diseases associated with such infections. They are in particular persistent chronic hepatitis, active chronic hepatitis, cirrhosis of the liver and hepatocarcinomas.  
         [0073]    A composition according to the invention may be administered by any conventional route used in the field of vaccines, in particular by the parenteral (e.g. subcutaneous, intradermal, intramuscular, intravenous or intraperitoneal) route. The choice of the route of administration depends on a number of parameters such as the nature of the active principle, polypeptide or DNA molecule, the adjuvant combined with the polypeptide or with the DNA molecule.  
         [0074]    A composition according to the invention may comprise, in addition to a polypeptide or a mixture of polypeptides useful for the purposes of the present invention, at least one other HCV antigen such as the E2 protein or alternatively such as a nonstructural protein NS1, NS2, NS3, NS4 or NS5, or a subunit, fragment, homolog, mutant or derivative of these antigens.  
         [0075]    A polypeptide, a mixture or a molecule of DNA useful for the purposes of the present invention may be formulated in or with liposomes, preferably neutral or anionic liposomes, microspheres ISCOMs or virus-like particles (VLPs), in order to promote the screening of the protein or of the polypeptide or to increase the immune response. Persons skilled in the art have these compounds available without difficulty; for example see Liposomes: A Practical Approach. RRC New Ed, IRL press (1990).  
         [0076]    Adjuvants other than liposomes may also be used. A large number are known to persons skilled in the art. Such adjuvants are identified by references below:  
         [0077]    For parenteral administration, there may be mentioned in particular aluminum compounds such as aluminum hydroxide, aluminum phosphate and aluminum hydroxyphosphate. The antigen may be absorbed or precipitated on an aluminum compound according to standard methods. Other adjuvants useful for parenteral administration include in particular polyphosphazene (WO 95/2415), DC-chol (3-beta-[N-(N′, N′-dimethylaminomethane) carbamoyl) cholesterol] (U.S. Pat. No. 5,283,185 and WO 96/14831), QS-21 (WO 88/9336) and RIBI from ImmunoChem (Hamilton, Mont.).  
         [0078]    The administration may take place in a single dose or in a dose repeated once or several times after a certain period. The appropriate dosage varies according to various parameters, for example the individual treated (adult or child), the vaccinal antigen itself, the mode and frequency of administration, the presence or absence of adjuvant and, if present, the type of adjuvant and the desired effect (e.g. protection or treatment), as will be determined by persons skilled in the art.  
         [0079]    A composition according to the invention may be manufactured conventionally. In particular, a polypeptide, a mixture or a molecule of DNA contained in the composition according to the invention is combined with a pharmaceutically acceptable diluent or carrier, e.g. water or a saline solution such as phosphate-buffered saline (PBS). In general, the diluent or the carrier is selected on the basis of the mode and route of administration and of standard pharmaceutical practices. Pharmaceutically acceptable diluents and carriers as well as all that is necessary for their use in pharmaceutical formulations are described in Remington&#39;s Pharmaceutical Sciences, a standard reference text in this field and in USP/NP.  
         [0080]    Exemplification  
       EXAMPLE 1  
     Construction of Recombinant Plasmids and Site-directed Mutagenesis  
       [0081]    The constructs called pRC/E1, pRC/CE1M1, pRC/CE1M2 and pRC/CE1M1M2 (also called pRC/CE1DM), pRC/C191M1 and pRC/C173, which are used below in Example 2, have been described in: Liu, Q., et al., J. Virol. 71:657 (1997). The inserts used are represented in FIG. 2. All the constructs are produced in the vector pRC which is obtained from InVitrogen (ref.: V780-20). The vector pRC carries the ampicillin gene and allows the expression of inserts under the control of a CMV promoter. Mutations called M1 and M2 are present in the constructs pRC/C 191 Ml, pRC/CE1M1, pRC/CE1M2 and pRC/CE1M1M2. They were generated by site-directed mutagenesis performed by PCR. The mutation called M1 corresponds to the replacement of the amino acids Serine 173  and Phe 174  of the C protein with the amino acids methionine and leucine, respectively. The mutation called M2 corresponds to the replacement of the amino acids alanine 191  and tyrosine 192  of the CE1 protein with the amino acids valine and asparagine, respectively.  
         [0082]    The plasmids expressing the reporter genes for luciferase and for P-galactosidase were constructed by modifying the vector pUC 18 (Appligene; ref: 161131). The expression of the genes is under the control of the immediate-early promoter 1 (ie1) of the human CMV. Sequences derived from the 3′ region of the bovine gene for the growth hormone were moreover added in 3′ of the genes in order to stabilize the mRNAs. These plasmids carry more than one ampicillin gene.  
       EXAMPLE 2  
     Transfection of Cells With the Plasmids  
       [0083]    CHO-K1 cells (ATCC CCL 61) were stored in a-MEM medium (Nature 230:310 (1971)), supplemented with 10% Foetal Calf Serum (FCS) (Hyclone, ref: A1115-L) and 20% Dimethyl Sulfoxide (DMSO) in liquid nitrogen. These cells are cultured under humid atmosphere at 37° C. with 5% CO 2  and 95% air. To carry out subcultures, the medium removed and the cellular lawn is rinsed with 5 ml of phosphate buffer (PBS). The supernatant is then removed before addition of 1.5 ml of trypsine per 75 cm 2  flask (trypsine at 0.025%). After incubating for 10 min in an incubator at 37° C., the reaction is stopped by addition of 10 ml of α-MEM medium containing 10% FCS. The cells are counted on a Malassez cell after a one-half dilution in 0.02% Trypan blue. 5×10 5  cells are then inoculated into dishes 6 cm in diameter with complete medium.  
         [0084]    The CHO cells are then cotransfected with one of the recombinant plasmids (PHCV) described avobe and a reporter plasmid (PCMV) which contains either the β-galactosidase gene under the control of the CMV promoter (pCMV β-gal) or the luciferase gene under the control of the CMV promoter (PCMV Luc).  
         [0085]    For that, 5 μg Of DNA (4.5 μg of plasmid pHCV/0.5 μg of plasmid pCMV) are diluted in 500 μl of OPTI-MEM medium (Gibco), and mixed with 14 μl of lipofectamine diluted in 500 μl of the same medium. The two solutions are mixed and incubated for 20 min at room temperature in order to allow the formation of the DNA-liposome complexes.  
         [0086]    The DNA liposome mixture diluted with 2 ml of OPTI-MEM is then added to the cells after removing the culture medium and rinsing in PBS. After incubating for 5 hours, the medium is again changed in 48 hr. after the transfection, it is then possible to test for the transient expression of the recombinant genes.  
       EXAMPLE 3  
     Demonstration of the Regulatory Activity of the Constructs on Reporter Genes  
       [0087]    The transfected cells are lysed with the aid of the reagent “Luciferase Cell Culture Lysis Reagent” (Promega, Luciferase Assay System). 100 μl of substrate are added to 100 μl of cell supernatant, directly by the bioluminometer injector (Lumat LB/9501/16 from Berthold) which measures the quantity of light emitted (Relative Light Units) for 10 seconds. The quantity of light emitted is then converted to nanograms of proteins per ml of cell lysate, by comparing with a standard curve established with the aid of purified luciferase.  
         [0088]    The results, which are presented in FIG. 2, show that a point mutation at the amino acid 191 in the construct CE1M2 abolishes the transactivating effect. A point mutation at the amino acid 173 (in the construct CE1M1) aabolishes the transactivating effect only in the case where C is fused with E1. A double mutation at the amino acids 173 and 191 abolishes the transactivating effect.  
     
       
       
         1 
         
           
             2  
           
           
             1  
             9379  
             DNA  
             Virus  
             
               CDS  
               (320)...(9352)  
             
           
            1 

cactccacca tgaatcactc ccctgtgagg aactactgtc ttcacgcaga aagcgtctag     60 

ccatggcgtt agtatgagtg tcgtgcagcc tccaggaccc cccctcccgg gagagccata    120 

gtggtctgcg gaaccggtga gtacaccgga attgccagga cgaccgggtc ctttcttgga    180 

tcaacccgct caatgcctgg agatttgggc gtgcccccgc aagactgcta gccgagtagt    240 

gttgggtcgc gaaaggcctt gtggtactgc ctgatagggt gcttgcgagt gccccgggag    300 

gtctcgtaga ccgtgcacc atg agc acg aat cct aaa cct caa aaa aaa aac     352 
                     Met Ser Thr Asn Pro Lys Pro Gln Lys Lys Asn 
                      1               5                   10 

aaa cgt aac acc aac cgt cgc cca cag gac gtc aag ttc ccg ggt ggc      400 
Lys Arg Asn Thr Asn Arg Arg Pro Gln Asp Val Lys Phe Pro Gly Gly 
             15                  20                  25 

ggt cag atc gtt ggt gga gtt tac ttg ttg ccg cgc agg ggc cct aga      448 
Gly Gln Ile Val Gly Gly Val Tyr Leu Leu Pro Arg Arg Gly Pro Arg 
         30                  35                  40 

ttg ggt gtg cgc gcg acg aga aag act tcc gag cgg tcg caa cct cga      496 
Leu Gly Val Arg Ala Thr Arg Lys Thr Ser Glu Arg Ser Gln Pro Arg 
     45                  50                  55 

ggt aga cgt cag cct atc ccc aag gct cgt cgg ccc gag ggc agg acc      544 
Gly Arg Arg Gln Pro Ile Pro Lys Ala Arg Arg Pro Glu Gly Arg Thr 
 60                  65                  70                  75 

tgg gct cag ccc ggg tac cct tgg ccc ctc tat ggc aat gag ggc tgc      592 
Trp Ala Gln Pro Gly Tyr Pro Trp Pro Leu Tyr Gly Asn Glu Gly Cys 
                 80                  85                  90 

ggg tgg gcg gga tgg ctc ctg tct ccc cgt ggc tct cgg cct agc tgg      640 
Gly Trp Ala Gly Trp Leu Leu Ser Pro Arg Gly Ser Arg Pro Ser Trp 
             95                 100                 105 

ggc ccc aca gac ccc cgg cgt agg tcg cgc aat ttg ggt aag gtc atc      688 
Gly Pro Thr Asp Pro Arg Arg Arg Ser Arg Asn Leu Gly Lys Val Ile 
        110                 115                 120 

gat acc ctt acg tgc ggc ttc gcc gac ctc atg ggg tac ata ccg ctc      736 
Asp Thr Leu Thr Cys Gly Phe Ala Asp Leu Met Gly Tyr Ile Pro Leu 
    125                 130                 135 

gtc ggc gcc cct ctt gga ggc gct gcc agg gcc ctg gcg cat ggc gtc      784 
Val Gly Ala Pro Leu Gly Gly Ala Ala Arg Ala Leu Ala His Gly Val 
140                 145                 150                 155 

cgg gtt ctg gaa gac ggc gtg aac tat gca aca ggg aac ctt cct ggt      832 
Arg Val Leu Glu Asp Gly Val Asn Tyr Ala Thr Gly Asn Leu Pro Gly 
                160                 165                 170 

tgc tct ttc tct atc ttc ctt ctg gcc ctg ctc tct tgc ttg act gtg      880 
Cys Ser Phe Ser Ile Phe Leu Leu Ala Leu Leu Ser Cys Leu Thr Val 
            175                 180                 185 

ccc gct tcg gcc tac caa gtg cgc aac tcc acg ggg ctt tac cac gtc      928 
Pro Ala Ser Ala Tyr Gln Val Arg Asn Ser Thr Gly Leu Tyr His Val 
        190                 195                 200 

acc aat gat tgc cct aac tcg agt att gtg tac gag gcg gcc gat gcc      976 
Thr Asn Asp Cys Pro Asn Ser Ser Ile Val Tyr Glu Ala Ala Asp Ala 
    205                 210                 215 

atc ctg cac act ccg ggg tgc gtc cct tgc gtt cgt gag ggc aac gcc     1024 
Ile Leu His Thr Pro Gly Cys Val Pro Cys Val Arg Glu Gly Asn Ala 
220                 225                 230                 235 

tcg agg tgt tgg gtg gcg atg acc cct acg gtg gcc acc agg gat ggc     1072 
Ser Arg Cys Trp Val Ala Met Thr Pro Thr Val Ala Thr Arg Asp Gly 
                240                 245                 250 

aaa ctc ccc gcg acg cag ctt cga cgt cac atc gat ctg ctt gtc ggg     1120 
Lys Leu Pro Ala Thr Gln Leu Arg Arg His Ile Asp Leu Leu Val Gly 
            255                 260                 265 

agc gcc acc ctc tgt tcg gcc ctc tac gtg ggg gac cta tgc ggg tct     1168 
Ser Ala Thr Leu Cys Ser Ala Leu Tyr Val Gly Asp Leu Cys Gly Ser 
        270                 275                 280 

gtc ttt ctt gtc ggc caa ctg ttc acc ttc tct ccc agg cgc cac tgg     1216 
Val Phe Leu Val Gly Gln Leu Phe Thr Phe Ser Pro Arg Arg His Trp 
    285                 290                 295 

acg acg caa ggt tgc aat tgc tct atc tat ccc ggc cat ata acg ggt     1264 
Thr Thr Gln Gly Cys Asn Cys Ser Ile Tyr Pro Gly His Ile Thr Gly 
300                 305                 310                 315 

cac cgc atg gca tgg gat atg atg atg aac tgg tcc cct acg acg gcg     1312 
His Arg Met Ala Trp Asp Met Met Met Asn Trp Ser Pro Thr Thr Ala 
                320                 325                 330 

ttg gta atg gct cag ctg ctc cgg atc cca caa gcc atc ttg gac atg     1360 
Leu Val Met Ala Gln Leu Leu Arg Ile Pro Gln Ala Ile Leu Asp Met 
            335                 340                 345 

atc gct ggt gct cac tgg gga gtc ctg gcg ggc ata gcg tat ttc tcc     1408 
Ile Ala Gly Ala His Trp Gly Val Leu Ala Gly Ile Ala Tyr Phe Ser 
        350                 355                 360 

atg gtg ggg aac tgg gcg aag gtc ctg gta gtg ctg ctg cta ttt gcc     1456 
Met Val Gly Asn Trp Ala Lys Val Leu Val Val Leu Leu Leu Phe Ala 
    365                 370                 375 

ggc gtc gac gcg gaa acc cac gtc acc ggg gga agt gcc ggc cac act     1504 
Gly Val Asp Ala Glu Thr His Val Thr Gly Gly Ser Ala Gly His Thr 
380                 385                 390                 395 

gtg tct gga ttt gtt agc ctc ctc gca cca ggc gcc aag cag aac gtc     1552 
Val Ser Gly Phe Val Ser Leu Leu Ala Pro Gly Ala Lys Gln Asn Val 
                400                 405                 410 

cag ctg atc aac acc aac ggc agt tgg cac ctc aat agc acg gcc ctg     1600 
Gln Leu Ile Asn Thr Asn Gly Ser Trp His Leu Asn Ser Thr Ala Leu 
            415                 420                 425 

aac tgc aat gat agc ctc aac acc ggc tgg ttg gca ggg ctt ttc tat     1648 
Asn Cys Asn Asp Ser Leu Asn Thr Gly Trp Leu Ala Gly Leu Phe Tyr 
        430                 435                 440 

cac cac aag ttc aac tct tca ggc tgt cct gag agg cta gcc agc tgc     1696 
His His Lys Phe Asn Ser Ser Gly Cys Pro Glu Arg Leu Ala Ser Cys 
    445                 450                 455 

cga ccc ctt acc gat ttt gac cag ggc tgg ggc cct atc agt tat gcc     1744 
Arg Pro Leu Thr Asp Phe Asp Gln Gly Trp Gly Pro Ile Ser Tyr Ala 
460                 465                 470                 475 

aac gga agc ggc ccc gac cag cgc ccc tac tgc tgg cac tac ccc cca     1792 
Asn Gly Ser Gly Pro Asp Gln Arg Pro Tyr Cys Trp His Tyr Pro Pro 
                480                 485                 490 

aaa cct tgc ggt att gtg ccc gcg aag agt gtg tgt ggt ccg gta tat     1840 
Lys Pro Cys Gly Ile Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr 
            495                 500                 505 

tgc ttc act ccc agc ccc gtg gtg gtg gga acg acc gac agg tcg ggc     1888 
Cys Phe Thr Pro Ser Pro Val Val Val Gly Thr Thr Asp Arg Ser Gly 
        510                 515                 520 

gcg ccc acc tac agc tgg ggt gaa aat gat acg gac gtc ttc gtc ctt     1936 
Ala Pro Thr Tyr Ser Trp Gly Glu Asn Asp Thr Asp Val Phe Val Leu 
    525                 530                 535 

aac aat acc agg cca ccg ctg ggc aat tgg ttc ggt tgt acc tgg atg     1984 
Asn Asn Thr Arg Pro Pro Leu Gly Asn Trp Phe Gly Cys Thr Trp Met 
540                 545                 550                 555 

aac tca act gga ttc acc aaa gtg tgc gga gcg cct cct tgt gtc atc     2032 
Asn Ser Thr Gly Phe Thr Lys Val Cys Gly Ala Pro Pro Cys Val Ile 
                560                 565                 570 

gga ggg gcg ggc aac aac acc ctg cac tgc ccc act gat tgc ttc cgc     2080 
Gly Gly Ala Gly Asn Asn Thr Leu His Cys Pro Thr Asp Cys Phe Arg 
            575                 580                 585 

aag cat ccg gac gcc aca tac tct cgg tgc ggc tcc ggt ccc tgg atc     2128 
Lys His Pro Asp Ala Thr Tyr Ser Arg Cys Gly Ser Gly Pro Trp Ile 
        590                 595                 600 

aca ccc agg tgc ctg gtc gac tac ccg tat agg ctt tgg cat tat cct     2176 
Thr Pro Arg Cys Leu Val Asp Tyr Pro Tyr Arg Leu Trp His Tyr Pro 
    605                 610                 615 

tgt acc atc aac tac acc ata ttt aaa atc agg atg tac gtg gga ggg     2224 
Cys Thr Ile Asn Tyr Thr Ile Phe Lys Ile Arg Met Tyr Val Gly Gly 
620                 625                 630                 635 

gtc gaa cac agg ctg gaa gct gcc tgc aac tgg acg cgg ggc gaa cgt     2272 
Val Glu His Arg Leu Glu Ala Ala Cys Asn Trp Thr Arg Gly Glu Arg 
                640                 645                 650 

tgc gat ctg gaa gac agg gac agg tcc gag ctc agc ccg tta ctg ctg     2320 
Cys Asp Leu Glu Asp Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu 
            655                 660                 665 

acc act aca cag tgg cag gtc ctc ccg tgt tcc ttc aca acc cta cca     2368 
Thr Thr Thr Gln Trp Gln Val Leu Pro Cys Ser Phe Thr Thr Leu Pro 
        670                 675                 680 

gcc ttg tcc acc ggc ctc atc cac ctc cac cag aac att gtg gac gtg     2416 
Ala Leu Ser Thr Gly Leu Ile His Leu His Gln Asn Ile Val Asp Val 
    685                 690                 695 

cag tac ttg tac ggg gtg ggg tca agc atc gcg tcc tgg gcc att aag     2464 
Gln Tyr Leu Tyr Gly Val Gly Ser Ser Ile Ala Ser Trp Ala Ile Lys 
700                 705                 710                 715 

tgg gag tac gtc gtt ctc ctg ttc ctt ctg ctt gca gac gcg cgc gtc     2512 
Trp Glu Tyr Val Val Leu Leu Phe Leu Leu Leu Ala Asp Ala Arg Val 
                720                 725                 730 

tgc tcc tgc ttg tgg atg atg cta ctc ata tcc caa gcg gag gcg gct     2560 
Cys Ser Cys Leu Trp Met Met Leu Leu Ile Ser Gln Ala Glu Ala Ala 
            735                 740                 745 

ttg gag aac ctc gta ata ctt aat gca gca tcc ctg gcc ggg acg cac     2608 
Leu Glu Asn Leu Val Ile Leu Asn Ala Ala Ser Leu Ala Gly Thr His 
        750                 755                 760 

ggt ctt gta tcc ttc ctc gtg ttc ttc tgc ttt gca tgg tat ttg aag     2656 
Gly Leu Val Ser Phe Leu Val Phe Phe Cys Phe Ala Trp Tyr Leu Lys 
    765                 770                 775 

ggt aag tgg gtg ccc gga gcg gtc tac acc ttc tac ggg atg tgg cct     2704 
Gly Lys Trp Val Pro Gly Ala Val Tyr Thr Phe Tyr Gly Met Trp Pro 
780                 785                 790                 795 

ctc ctc ctg ctc ctg ttg gcg ttg ccc cag cgg gcg tac gcg ctg gac     2752 
Leu Leu Leu Leu Leu Leu Ala Leu Pro Gln Arg Ala Tyr Ala Leu Asp 
                800                 805                 810 

acg gag gtg gcc gcg tcg tgt ggc ggt gtt gtt ctc gtc ggg ttg atg     2800 
Thr Glu Val Ala Ala Ser Cys Gly Gly Val Val Leu Val Gly Leu Met 
            815                 820                 825 

gcg ctg act ctg tca cca tat tac aag cgc tat atc agc tgg tgc ttg     2848 
Ala Leu Thr Leu Ser Pro Tyr Tyr Lys Arg Tyr Ile Ser Trp Cys Leu 
        830                 835                 840 

tgg tgg ctt cag tat ttt ctg acc aga gtg gaa gcg caa ctg cac gtg     2896 
Trp Trp Leu Gln Tyr Phe Leu Thr Arg Val Glu Ala Gln Leu His Val 
    845                 850                 855 

tgg att ccc ccc ctc aac gtc cga ggg ggg cgc gac gcc gtc atc tta     2944 
Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val Ile Leu 
860                 865                 870                 875 

ctc atg tgt gct gta cac ccg act ctg gta ttt gac atc acc aaa ttg     2992 
Leu Met Cys Ala Val His Pro Thr Leu Val Phe Asp Ile Thr Lys Leu 
                880                 885                 890 

ctg ctg gcc gtc ttc gga ccc ctt tgg att ctt caa gcc agt ttg ctt     3040 
Leu Leu Ala Val Phe Gly Pro Leu Trp Ile Leu Gln Ala Ser Leu Leu 
            895                 900                 905 

aaa gta ccc tac ttt gtg cgc gtc caa ggc ctt ctc cgg ttc tgc gcg     3088 
Lys Val Pro Tyr Phe Val Arg Val Gln Gly Leu Leu Arg Phe Cys Ala 
        910                 915                 920 

tta gcg cgg aag atg atc gga ggc cat tac gtg caa atg gtc atc att     3136 
Leu Ala Arg Lys Met Ile Gly Gly His Tyr Val Gln Met Val Ile Ile 
    925                 930                 935 

aag tta ggg gcg ctt act ggc acc tat gtt tat aac cat ctc act cct     3184 
Lys Leu Gly Ala Leu Thr Gly Thr Tyr Val Tyr Asn His Leu Thr Pro 
940                 945                 950                 955 

ctt cgg gac tgg gcg cac aac ggc ttg cga gat ctg gcc gtg gct gta     3232 
Leu Arg Asp Trp Ala His Asn Gly Leu Arg Asp Leu Ala Val Ala Val 
                960                 965                 970 

gag cca gtc gtc ttc tcc caa atg gag acc aag ctc atc acg tgg ggg     3280 
Glu Pro Val Val Phe Ser Gln Met Glu Thr Lys Leu Ile Thr Trp Gly 
            975                 980                 985 

gca gat acc gcc gcg tgc ggt gac atc atc aac ggc ttg cct gtt tcc     3328 
Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Asn Gly Leu Pro Val Ser 
         990                 995                1000 

gcc cgc agg ggc cgg gag ata ctg ctc ggg cca gcc gat gga atg gtc     3376 
Ala Arg Arg Gly Arg Glu Ile Leu Leu Gly Pro Ala Asp Gly Met Val 
    1005                1010                1015 

tcc aag ggg tgg agg ttg ctg gcg ccc atc acg gcg tac gcc cag cag     3424 
Ser Lys Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr Ala Gln Gln 
1020                1025                1030                1035 

aca agg ggc ctc cta ggg tgc ata atc acc agc cta act ggc cgg gac     3472 
Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp 
                1040                1045                1050 

aaa aac caa gtg gag ggt gag gtc cag att gtg tca act gct gcc caa     3520 
Lys Asn Gln Val Glu Gly Glu Val Gln Ile Val Ser Thr Ala Ala Gln 
            1055                1060                1065 

acc ttc ctg gca acg tgc atc aat ggg gtg tgc tgg act gtc tac cac     3568 
Thr Phe Leu Ala Thr Cys Ile Asn Gly Val Cys Trp Thr Val Tyr His 
        1070                1075                1080 

ggg gcc gga acg agg acc atc gcg tca ccc aag ggt cct gtc atc cag     3616 
Gly Ala Gly Thr Arg Thr Ile Ala Ser Pro Lys Gly Pro Val Ile Gln 
    1085                1090                1095 

atg tat acc aat gta gac caa gac ctt gtg ggc tgg ccc gct ccg caa     3664 
Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly Trp Pro Ala Pro Gln 
1100                1105                1110                1115 

ggt agc cgc tca ttg aca ccc tgc act tgc ggc tcc tcg gac ctt tac     3712 
Gly Ser Arg Ser Leu Thr Pro Cys Thr Cys Gly Ser Ser Asp Leu Tyr 
                1120                1125                1130 

ctg gtc acg agg cac gcc gat gtc att ccc gtg cgc cgg cgg ggt gat     3760 
Leu Val Thr Arg His Ala Asp Val Ile Pro Val Arg Arg Arg Gly Asp 
            1135                1140                1145 

agc agg ggc agc ctg ctg tcg ccc cgg ccc att tcc tac ttg aaa ggc     3808 
Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro Ile Ser Tyr Leu Lys Gly 
        1150                1155                1160 

tcc tcg ggg ggt ccg ctg ttg tgc ccc gcg ggg cac gcc gtg ggc ata     3856 
Ser Ser Gly Gly Pro Leu Leu Cys Pro Ala Gly His Ala Val Gly Ile 
    1165                1170                1175 

ttt agg gcc gcg gtg tgc acc cgt gga gtg gct aag gcg gtg gac ttt     3904 
Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala Lys Ala Val Asp Phe 
1180                1185                1190                1195 

atc cct gtg gag aac cta gag aca acc atg agg tcc ccg gtg ttc acg     3952 
Ile Pro Val Glu Asn Leu Glu Thr Thr Met Arg Ser Pro Val Phe Thr 
                1200                1205                1210 

gat aac tcc tct cca cca gta gtg ccc cag agc ttc cag gtg gct cac     4000 
Asp Asn Ser Ser Pro Pro Val Val Pro Gln Ser Phe Gln Val Ala His 
            1215                1220                1225 

ctc cat gct ccc aca ggc agc ggc aaa agc acc aag gtc ccg gct gca     4048 
Leu His Ala Pro Thr Gly Ser Gly Lys Ser Thr Lys Val Pro Ala Ala 
        1230                1235                1240 

tat gca gct cag ggc tat aag gtg cta gta ctc aac ccc tct gtt gct     4096 
Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala 
    1245                1250                1255 

gca aca ctg ggc ttt ggt gct tac atg tcc aag gct cat ggg atc gat     4144 
Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly Ile Asp 
1260                1265                1270                1275 

cct aac atc agg acc ggg gtg aga aca att acc act ggc agc ccc atc     4192 
Pro Asn Ile Arg Thr Gly Val Arg Thr Ile Thr Thr Gly Ser Pro Ile 
                1280                1285                1290 

acg tac tcc acc tac ggc aag ttc ctt gcc gac ggc ggg tgc tcg ggg     4240 
Thr Tyr Ser Thr Tyr Gly Lys Phe Leu Ala Asp Gly Gly Cys Ser Gly 
            1295                1300                1305 

ggc gct tat gac ata ata att tgt gac gag tgc cac tcc acg gat gcc     4288 
Gly Ala Tyr Asp Ile Ile Ile Cys Asp Glu Cys His Ser Thr Asp Ala 
        1310                1315                1320 

aca tcc atc ttg ggc atc ggc act gtc ctt gac caa gca gag act gcg     4336 
Thr Ser Ile Leu Gly Ile Gly Thr Val Leu Asp Gln Ala Glu Thr Ala 
    1325                1330                1335 

ggg gcg aga ctg gtt gtg ctc gcc acc gcc acc cct ccg ggc tcc gtc     4384 
Gly Ala Arg Leu Val Val Leu Ala Thr Ala Thr Pro Pro Gly Ser Val 
1340                1345                1350                1355 

act gtg ccc cat ccc aac atc gag gag gtt gct ctg tcc acc acc gga     4432 
Thr Val Pro His Pro Asn Ile Glu Glu Val Ala Leu Ser Thr Thr Gly 
                1360                1365                1370 

gag atc cct ttt tac ggc aag gct atc ccc ctc gaa gta atc aag ggg     4480 
Glu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Leu Glu Val Ile Lys Gly 
            1375                1380                1385 

ggg aga cat ctc atc ttc tgt cat tca aag aag aag tgc gac gaa ctc     4528 
Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys Asp Glu Leu 
        1390                1395                1400 

gcc gca aag ctg gtc gca ttg ggc atc aat gcc gtg gcc tac tac cgc     4576 
Ala Ala Lys Leu Val Ala Leu Gly Ile Asn Ala Val Ala Tyr Tyr Arg 
    1405                1410                1415 

ggt ctt gac gtg tcc gtc atc ccg acc agc ggc gat gtt gtc gtc gtg     4624 
Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Val Val Val 
1420                1425                1430                1435 

gca acc gat gcc ctc atg acc ggc tat acc ggc gac ttc gac tcg gtg     4672 
Ala Thr Asp Ala Leu Met Thr Gly Tyr Thr Gly Asp Phe Asp Ser Val 
                1440                1445                1450 

ata gac tgc aat acg tgt gtc acc cag aca gtc gat ttc agc ctt gac     4720 
Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val Asp Phe Ser Leu Asp 
            1455                1460                1465 

cct acc ttc acc att gag aca atc acg ctc ccc cag gat gct gtc tcc     4768 
Pro Thr Phe Thr Ile Glu Thr Ile Thr Leu Pro Gln Asp Ala Val Ser 
        1470                1475                1480 

cgc act caa cgt cgg ggc agg act ggc agg ggg aag cca ggc atc tac     4816 
Arg Thr Gln Arg Arg Gly Arg Thr Gly Arg Gly Lys Pro Gly Ile Tyr 
    1485                1490                1495 

aga ttt gtg gca ccg ggg gag cgc ccc tcc ggc atg ttc gac tcg tcc     4864 
Arg Phe Val Ala Pro Gly Glu Arg Pro Ser Gly Met Phe Asp Ser Ser 
1500                1505                1510                1515 

gtc ctc tgt gag tgc tat gac gca ggc tgt gct tgg tat gag ctc acg     4912 
Val Leu Cys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr Glu Leu Thr 
                1520                1525                1530 

ccc gcc gag act aca gtt agg cta cga gcg tac atg aac acc ccg ggg     4960 
Pro Ala Glu Thr Thr Val Arg Leu Arg Ala Tyr Met Asn Thr Pro Gly 
            1535                1540                1545 

ctt ccc gtg tgc cag gac cat ctt gaa ttt tgg gag ggc gtc ttt aca     5008 
Leu Pro Val Cys Gln Asp His Leu Glu Phe Trp Glu Gly Val Phe Thr 
        1550                1555                1560 

ggc ctc act cat ata gat gcc cac ttt cta tcc cag aca aag cag agt     5056 
Gly Leu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr Lys Gln Ser 
    1565                1570                1575 

ggg gag aac ctt cct tac ctg gta gcg tac caa gcc acc gtg tgc gct     5104 
Gly Glu Asn Leu Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val Cys Ala 
1580                1585                1590                1595 

agg gct caa gcc cct ccc cca tcg tgg gac cag atg tgg aag tgt ttg     5152 
Arg Ala Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp Lys Cys Leu 
                1600                1605                1610 

att cgc ctc aag ccc acc ctc cat ggg cca aca ccc ctg cta tac aga     5200 
Ile Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu Tyr Arg 
            1615                1620                1625 

ctg ggc gct gtt cag aat gaa atc acc ctg acg cac cca gtc acc aaa     5248 
Leu Gly Ala Val Gln Asn Glu Ile Thr Leu Thr His Pro Val Thr Lys 
        1630                1635                1640 

tac atc atg aca tgc atg tcg gcc gac ctg gag gtc gtc acg agc acc     5296 
Tyr Ile Met Thr Cys Met Ser Ala Asp Leu Glu Val Val Thr Ser Thr 
    1645                1650                1655 

tgg gtg ctc gtt ggc ggc gtc ctg gct gct ttg gcc gcg tat tgc ctg     5344 
Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr Cys Leu 
1660                1665                1670                1675 

tca aca ggc tgc gtg gtc ata gtg ggc agg gtc gtc ttg tcc ggg aag     5392 
Ser Thr Gly Cys Val Val Ile Val Gly Arg Val Val Leu Ser Gly Lys 
                1680                1685                1690 

ccg gca atc ata cct gac agg gaa gtc ctc tac cga gag ttc gat gag     5440 
Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu Phe Asp Glu 
            1695                1700                1705 

atg gaa gag tgc tct cag cac tta ccg tac atc gag caa ggg atg atg     5488 
Met Glu Glu Cys Ser Gln His Leu Pro Tyr Ile Glu Gln Gly Met Met 
        1710                1715                1720 

ctc gcc gag cag ttc aag cag aag gcc ctc ggc ctc ctg cag acc gcg     5536 
Leu Ala Glu Gln Phe Lys Gln Lys Ala Leu Gly Leu Leu Gln Thr Ala 
    1725                1730                1735 

tcc cgt cag gca gag gtt atc gcc cct gct gtc cag acc aac tgg caa     5584 
Ser Arg Gln Ala Glu Val Ile Ala Pro Ala Val Gln Thr Asn Trp Gln 
1740                1745                1750                1755 

aaa ctc gag acc ttc tgg gcg aag cat atg tgg aac ttc atc agt ggg     5632 
Lys Leu Glu Thr Phe Trp Ala Lys His Met Trp Asn Phe Ile Ser Gly 
                1760                1765                1770 

ata caa tac ttg gcg ggc ttg tca acg ctg cct ggt aac ccc gcc att     5680 
Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro Ala Ile 
            1775                1780                1785 

gct tca ttg atg gct ttt aca gct gct gtc acc agc cca cta acc act     5728 
Ala Ser Leu Met Ala Phe Thr Ala Ala Val Thr Ser Pro Leu Thr Thr 
        1790                1795                1800 

agc caa acc ctc ctc ttc aac ata ttg ggg ggg tgg gtg gct gcc cag     5776 
Ser Gln Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala Ala Gln 
    1805                1810                1815 

ctc gcc gcc ccc ggt gcc gct act gcc ttt gtg ggc gct ggc tta gct     5824 
Leu Ala Ala Pro Gly Ala Ala Thr Ala Phe Val Gly Ala Gly Leu Ala 
1820                1825                1830                1835 

ggc gcc gcc atc ggc agt gtt gga ctg ggg aag gtc ctc ata gac atc     5872 
Gly Ala Ala Ile Gly Ser Val Gly Leu Gly Lys Val Leu Ile Asp Ile 
                1840                1845                1850 

ctt gca ggg tat ggc gcg ggc gtg gcg gga gct ctt gtg gca ttc aag     5920 
Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala Phe Lys 
            1855                1860                1865 

atc atg agc ggt gag gtc ccc tcc acg gag gac ctg gtc aat cta ctg     5968 
Ile Met Ser Gly Glu Val Pro Ser Thr Glu Asp Leu Val Asn Leu Leu 
        1870                1875                1880 

ccc gcc atc ctc tcg ccc gga gcc ctc gta gtc ggc gtg gtc tgt gca     6016 
Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly Val Val Cys Ala 
    1885                1890                1895 

gca ata ctg cgc cgg cac gtt ggc ccg ggc gag ggg gca gtg cag tgg     6064 
Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu Gly Ala Val Gln Trp 
1900                1905                1910                1915 

atg aac cgg ctg ata gcc ttc gcc tcc cgg ggg aac cat gtt tcc ccc     6112 
Met Asn Arg Leu Ile Ala Phe Ala Ser Arg Gly Asn His Val Ser Pro 
                1920                1925                1930 

acg cac tac gtg ccg gag agc gat gca gct gcc cgc gtc act gcc ata     6160 
Thr His Tyr Val Pro Glu Ser Asp Ala Ala Ala Arg Val Thr Ala Ile 
            1935                1940                1945 

ctc agc agc ctc act gta acc cag ctc ctg agg cga ctg cac cag tgg     6208 
Leu Ser Ser Leu Thr Val Thr Gln Leu Leu Arg Arg Leu His Gln Trp 
        1950                1955                1960 

ata agc tcg gag tgt acc act cca tgc tcc ggt tcc tgg cta agg gac     6256 
Ile Ser Ser Glu Cys Thr Thr Pro Cys Ser Gly Ser Trp Leu Arg Asp 
    1965                1970                1975 

atc tgg gac tgg ata tgc gag gtg ttg agc gac ttt aag acc tgg cta     6304 
Ile Trp Asp Trp Ile Cys Glu Val Leu Ser Asp Phe Lys Thr Trp Leu 
1980                1985                1990                1995 

aaa gct aag ctc atg cca cag ctg cct ggg atc ccc ttt gtg tcc tgc     6352 
Lys Ala Lys Leu Met Pro Gln Leu Pro Gly Ile Pro Phe Val Ser Cys 
                2000                2005                2010 

cag cgc ggg tat aag ggg gtc tgg cga gtg gac ggc atc atg cac act     6400 
Gln Arg Gly Tyr Lys Gly Val Trp Arg Val Asp Gly Ile Met His Thr 
            2015                2020                2025 

cgc tgc cac tgt gga gct gag atc act gga cat gtc aaa aac ggg acg     6448 
Arg Cys His Cys Gly Ala Glu Ile Thr Gly His Val Lys Asn Gly Thr 
        2030                2035                2040 

atg agg atc gtc ggt cct agg acc tgc agg aac atg tgg agt ggg acc     6496 
Met Arg Ile Val Gly Pro Arg Thr Cys Arg Asn Met Trp Ser Gly Thr 
    2045                2050                2055 

ttc ccc att aat gcc tac acc acg ggc ccc tgt acc ccc ctt cct gcg     6544 
Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Leu Pro Ala 
2060                2065                2070                2075 

ccg aac tac acg ttc gcg cta tgg agg gtg tct gca gag gaa tat gtg     6592 
Pro Asn Tyr Thr Phe Ala Leu Trp Arg Val Ser Ala Glu Glu Tyr Val 
                2080                2085                2090 

gag ata agg cag gtg ggg gac ttc cac tac gtg acg ggt atg act act     6640 
Glu Ile Arg Gln Val Gly Asp Phe His Tyr Val Thr Gly Met Thr Thr 
            2095                2100                2105 

gac aat ctc aaa tgc ccg tgc cag gtc cca tcg ccc gaa ttt ttc aca     6688 
Asp Asn Leu Lys Cys Pro Cys Gln Val Pro Ser Pro Glu Phe Phe Thr 
        2110                2115                2120 

gaa ttg gac ggg gtg cgc cta cat agg ttt gcg ccc ccc tgc aag ccc     6736 
Glu Leu Asp Gly Val Arg Leu His Arg Phe Ala Pro Pro Cys Lys Pro 
    2125                2130                2135 

ttg ctg cgg gag gag gta tca ttc aga gta gga ctc cac gaa tac ccg     6784 
Leu Leu Arg Glu Glu Val Ser Phe Arg Val Gly Leu His Glu Tyr Pro 
2140                2145                2150                2155 

gta ggg tcg caa tta cct tgc gag ccc gaa ccg gac gtg gcc gtg ttg     6832 
Val Gly Ser Gln Leu Pro Cys Glu Pro Glu Pro Asp Val Ala Val Leu 
                2160                2165                2170 

acg tcc atg ctc act gat ccc tcc cat ata aca gca gag gcg gcc ggg     6880 
Thr Ser Met Leu Thr Asp Pro Ser His Ile Thr Ala Glu Ala Ala Gly 
            2175                2180                2185 

cga agg ttg gcg agg gga tca ccc ccc tct gtg gcc agc tcc tcg gct     6928 
Arg Arg Leu Ala Arg Gly Ser Pro Pro Ser Val Ala Ser Ser Ser Ala 
        2190                2195                2200 

agc cag cta tcc gct cca tct ctc aag gca act tgc acc gct aac cat     6976 
Ser Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Ala Asn His 
    2205                2210                2215 

gac tcc cct gat gct gag ctc ata gag gcc aac ctc cta tgg agg cag     7024 
Asp Ser Pro Asp Ala Glu Leu Ile Glu Ala Asn Leu Leu Trp Arg Gln 
2220                2225                2230                2235 

gag atg ggc ggc aac atc acc agg gtt gag tca gaa aac aaa gtg gtg     7072 
Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn Lys Val Val 
                2240                2245                2250 

att ctg gac tcc ttc gat ccg ctt gtg gcg gag gag gac gag cgg gag     7120 
Ile Leu Asp Ser Phe Asp Pro Leu Val Ala Glu Glu Asp Glu Arg Glu 
            2255                2260                2265 

atc tcc gta ccc gca gaa atc ctg cgg aag tct cgg aga ttc gcc cag     7168 
Ile Ser Val Pro Ala Glu Ile Leu Arg Lys Ser Arg Arg Phe Ala Gln 
        2270                2275                2280 

gcc ctg ccc gtt tgg gcg cgg ccg gac tat aac ccc ccg cta gtg gag     7216 
Ala Leu Pro Val Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu Val Glu 
    2285                2290                2295 

acg tgg aaa aag ccc gac tac gaa cca cct gtg gtc cat ggc tgt ccg     7264 
Thr Trp Lys Lys Pro Asp Tyr Glu Pro Pro Val Val His Gly Cys Pro 
2300                2305                2310                2315 

ctt cca cct cca aag tcc cct cct gtg cct ccg cct cgg aag aag cgg     7312 
Leu Pro Pro Pro Lys Ser Pro Pro Val Pro Pro Pro Arg Lys Lys Arg 
                2320                2325                2330 

acg gtg gtc ctc act gaa tca acc cta tct act gcc ttg gcc gag ctc     7360 
Thr Val Val Leu Thr Glu Ser Thr Leu Ser Thr Ala Leu Ala Glu Leu 
            2335                2340                2345 

gcc acc aga agc ttt ggc agc tcc tca act tcc ggc att acg ggc gac     7408 
Ala Thr Arg Ser Phe Gly Ser Ser Ser Thr Ser Gly Ile Thr Gly Asp 
        2350                2355                2360 

aat acg aca aca tcc tct gag ccc gcc cct tct ggc tgc ccc ccc gac     7456 
Asn Thr Thr Thr Ser Ser Glu Pro Ala Pro Ser Gly Cys Pro Pro Asp 
    2365                2370                2375 

tcc gac gct gag tcc tat tcc tcc atg ccc ccc ctg gag ggg gag cct     7504 
Ser Asp Ala Glu Ser Tyr Ser Ser Met Pro Pro Leu Glu Gly Glu Pro 
2380                2385                2390                2395 

ggg gat ccg gat ctt agc gac ggg tca tgg tca acg gtc agt agt gag     7552 
Gly Asp Pro Asp Leu Ser Asp Gly Ser Trp Ser Thr Val Ser Ser Glu 
                2400                2405                2410 

gcc aac gcg gag gat gtc gtg tgc tgc tca atg tct tac tct tgg aca     7600 
Ala Asn Ala Glu Asp Val Val Cys Cys Ser Met Ser Tyr Ser Trp Thr 
            2415                2420                2425 

ggc gca ctc gtc acc ccg tgc gcc gcg gaa gaa cag aaa ctg ccc atc     7648 
Gly Ala Leu Val Thr Pro Cys Ala Ala Glu Glu Gln Lys Leu Pro Ile 
        2430                2435                2440 

aat gca cta agc aac tcg ttg cta cgt cac cac aat ttg gtg tat tcc     7696 
Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val Tyr Ser 
    2445                2450                2455 

acc acc tca cgc agt gct tgc caa agg cag aag aaa gtc aca ttt gac     7744 
Thr Thr Ser Arg Ser Ala Cys Gln Arg Gln Lys Lys Val Thr Phe Asp 
2460                2465                2470                2475 

aga ctg caa gtt ctg gac agc cat tac cag gac gta ctc aag gag gtt     7792 
Arg Leu Gln Val Leu Asp Ser His Tyr Gln Asp Val Leu Lys Glu Val 
                2480                2485                2490 

aaa gca gcg gcg tca aaa gtg aag gct aac ttg cta tcc gta gag gaa     7840 
Lys Ala Ala Ala Ser Lys Val Lys Ala Asn Leu Leu Ser Val Glu Glu 
            2495                2500                2505 

gct tgc agc ctg acg ccc cca cac tca gcc aaa tcc aag ttt ggt tat     7888 
Ala Cys Ser Leu Thr Pro Pro His Ser Ala Lys Ser Lys Phe Gly Tyr 
        2510                2515                2520 

ggg gca aaa gac gtc cgt tgc cat gcc aga aag gcc gta acc cac atc     7936 
Gly Ala Lys Asp Val Arg Cys His Ala Arg Lys Ala Val Thr His Ile 
    2525                2530                2535 

aac tcc gtg tgg aaa gac ctt ctg gaa gac aat gta aca cca ata gac     7984 
Asn Ser Val Trp Lys Asp Leu Leu Glu Asp Asn Val Thr Pro Ile Asp 
2540                2545                2550                2555 

act acc atc atg gct aag aac gag gtt ttc tgc gtt cag cct gag aag     8032 
Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val Gln Pro Glu Lys 
                2560                2565                2570 

ggg ggt cgt aag cca gct cgt ctc atc gtg ttc ccc gat ctg ggc gtg     8080 
Gly Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly Val 
            2575                2580                2585 

cgc gtg tgc gaa aag atg gct ttg tac gac gtg gtt aca aag ctc ccc     8128 
Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Thr Lys Leu Pro 
        2590                2595                2600 

ttg gcc gtg atg gga agc tcc tac gga ttc caa tac tca cca gga cag     8176 
Leu Ala Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly Gln 
    2605                2610                2615 

cgg gtt gaa ttc ctc gtg caa gcg tgg aag tcc aag aaa acc cca atg     8224 
Arg Val Glu Phe Leu Val Gln Ala Trp Lys Ser Lys Lys Thr Pro Met 
2620                2625                2630                2635 

ggg ttc tcg tat gat acc cgc tgc ttt gac tcc aca gtc act gag agc     8272 
Gly Phe Ser Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr Glu Ser 
                2640                2645                2650 

gac atc cgt acg gag gag gca atc tac caa tgt tgt gac ctc gac ccc     8320 
Asp Ile Arg Thr Glu Glu Ala Ile Tyr Gln Cys Cys Asp Leu Asp Pro 
            2655                2660                2665 

caa gcc cgc gtg gcc atc aag tcc ctc acc gag agg ctt tat gtt ggg     8368 
Gln Ala Arg Val Ala Ile Lys Ser Leu Thr Glu Arg Leu Tyr Val Gly 
        2670                2675                2680 

ggc cct ctt acc aat tca agg ggg gag aac tgc ggc tat cgc agg tgc     8416 
Gly Pro Leu Thr Asn Ser Arg Gly Glu Asn Cys Gly Tyr Arg Arg Cys 
    2685                2690                2695 

cgc gcg agc ggc gta ctg aca act agc tgt ggt aac acc ctc act tgc     8464 
Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr Cys 
2700                2705                2710                2715 

tac atc aag gcc cgg gca gcc tgt cga gcc gca ggg ctc cag gac tgc     8512 
Tyr Ile Lys Ala Arg Ala Ala Cys Arg Ala Ala Gly Leu Gln Asp Cys 
                2720                2725                2730 

acc atg ctc gtg tgt ggc gac gac tta gtc gtt atc tgt gaa agc gcg     8560 
Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser Ala 
            2735                2740                2745 

ggg gtc cag gag gac gcg gcg agc ctg aga gcc ttc acg gag gct atg     8608 
Gly Val Gln Glu Asp Ala Ala Ser Leu Arg Ala Phe Thr Glu Ala Met 
        2750                2755                2760 

acc agg tac tcc gcc ccc cct ggg gac ccc cca caa cca gaa tac gac     8656 
Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Gln Pro Glu Tyr Asp 
    2765                2770                2775 

ttg gag ctc ata aca tca tgc tcc tcc aac gtg tca gtc gcc cac gac     8704 
Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val Ser Val Ala His Asp 
2780                2785                2790                2795 

ggc gct gga aag agg gtc tac tac ctc acc cgt gac cct aca acc ccc     8752 
Gly Ala Gly Lys Arg Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr Pro 
                2800                2805                2810 

ctc gcg aga gct gcg tgg gag aca gca aga cac act cca gtc aat tcc     8800 
Leu Ala Arg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val Asn Ser 
            2815                2820                2825 

tgg cta ggc aac ata atc atg ttt gcc ccc aca ctg tgg gcg agg atg     8848 
Trp Leu Gly Asn Ile Ile Met Phe Ala Pro Thr Leu Trp Ala Arg Met 
        2830                2835                2840 

ata ctg atg acc cat ttc ttt agc gtc ctt ata gcc agg gac cag ctt     8896 
Ile Leu Met Thr His Phe Phe Ser Val Leu Ile Ala Arg Asp Gln Leu 
    2845                2850                2855 

gaa cag gcc ctc gat tgc gag atc tac ggg gcc tgc tac tcc ata gaa     8944 
Glu Gln Ala Leu Asp Cys Glu Ile Tyr Gly Ala Cys Tyr Ser Ile Glu 
2860                2865                2870                2875 

cca ctt gat cta cct cca atc att caa aga ctc cat ggc ctc agc gca     8992 
Pro Leu Asp Leu Pro Pro Ile Ile Gln Arg Leu His Gly Leu Ser Ala 
                2880                2885                2890 

ttt tca ctc cac agt tac tct cca ggt gaa att aat agg gtg gcc gca     9040 
Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala Ala 
            2895                2900                2905 

tgc ctc aga aaa ctt ggg gta ccg ccc ttg cga gct tgg aga cac cgg     9088 
Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg Ala Trp Arg His Arg 
        2910                2915                2920 

gcc cgg agc gtc cgc gct agg ctt ctg gcc aga gga ggc agg gct gcc     9136 
Ala Arg Ser Val Arg Ala Arg Leu Leu Ala Arg Gly Gly Arg Ala Ala 
    2925                2930                2935 

ata tgt ggc aag tac ctc ttc aac tgg gca gta aga aca aag ctc aaa     9184 
Ile Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu Lys 
2940                2945                2950                2955 

ctc act cca ata gcg gcc gct ggc cag ctg gac ttg tcc ggc tgg ttc     9232 
Leu Thr Pro Ile Ala Ala Ala Gly Gln Leu Asp Leu Ser Gly Trp Phe 
                2960                2965                2970 

acg gct ggc tac agc ggg gga gac att tat cac agc gtg tct cat gcc     9280 
Thr Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser Val Ser His Ala 
            2975                2980                2985 

cgg ccc cgc tgg atc tgg ttt tgc cta ctc ctg ctt gct gca ggg gta     9328 
Arg Pro Arg Trp Ile Trp Phe Cys Leu Leu Leu Leu Ala Ala Gly Val 
        2990                2995                3000 

ggc atc tac ctc ctc ccc aac cga tgaaggttgg ggtaaacact ccggcct       9379 
Gly Ile Tyr Leu Leu Pro Asn Arg 
    3005                3010 

 
           
             2  
             3011  
             PRT  
             Virus  
           
            2 

Met Ser Thr Asn Pro Lys Pro Gln Lys Lys Asn Lys Arg Asn Thr Asn 
 1               5                  10                  15 

Arg Arg Pro Gln Asp Val Lys Phe Pro Gly Gly Gly Gln Ile Val Gly 
            20                  25                  30 

Gly Val Tyr Leu Leu Pro Arg Arg Gly Pro Arg Leu Gly Val Arg Ala 
        35                  40                  45 

Thr Arg Lys Thr Ser Glu Arg Ser Gln Pro Arg Gly Arg Arg Gln Pro 
    50                  55                  60 

Ile Pro Lys Ala Arg Arg Pro Glu Gly Arg Thr Trp Ala Gln Pro Gly 
65                  70                  75                  80 

Tyr Pro Trp Pro Leu Tyr Gly Asn Glu Gly Cys Gly Trp Ala Gly Trp 
                85                  90                  95 

Leu Leu Ser Pro Arg Gly Ser Arg Pro Ser Trp Gly Pro Thr Asp Pro 
            100                 105                 110 

Arg Arg Arg Ser Arg Asn Leu Gly Lys Val Ile Asp Thr Leu Thr Cys 
        115                 120                 125 

Gly Phe Ala Asp Leu Met Gly Tyr Ile Pro Leu Val Gly Ala Pro Leu 
    130                 135                 140 

Gly Gly Ala Ala Arg Ala Leu Ala His Gly Val Arg Val Leu Glu Asp 
145                 150                 155                 160 

Gly Val Asn Tyr Ala Thr Gly Asn Leu Pro Gly Cys Ser Phe Ser Ile 
                165                 170                 175 

Phe Leu Leu Ala Leu Leu Ser Cys Leu Thr Val Pro Ala Ser Ala Tyr 
            180                 185                 190 

Gln Val Arg Asn Ser Thr Gly Leu Tyr His Val Thr Asn Asp Cys Pro 
        195                 200                 205 

Asn Ser Ser Ile Val Tyr Glu Ala Ala Asp Ala Ile Leu His Thr Pro 
    210                 215                 220 

Gly Cys Val Pro Cys Val Arg Glu Gly Asn Ala Ser Arg Cys Trp Val 
225                 230                 235                 240 

Ala Met Thr Pro Thr Val Ala Thr Arg Asp Gly Lys Leu Pro Ala Thr 
                245                 250                 255 

Gln Leu Arg Arg His Ile Asp Leu Leu Val Gly Ser Ala Thr Leu Cys 
            260                 265                 270 

Ser Ala Leu Tyr Val Gly Asp Leu Cys Gly Ser Val Phe Leu Val Gly 
        275                 280                 285 

Gln Leu Phe Thr Phe Ser Pro Arg Arg His Trp Thr Thr Gln Gly Cys 
    290                 295                 300 

Asn Cys Ser Ile Tyr Pro Gly His Ile Thr Gly His Arg Met Ala Trp 
305                 310                 315                 320 

Asp Met Met Met Asn Trp Ser Pro Thr Thr Ala Leu Val Met Ala Gln 
                325                 330                 335 

Leu Leu Arg Ile Pro Gln Ala Ile Leu Asp Met Ile Ala Gly Ala His 
            340                 345                 350 

Trp Gly Val Leu Ala Gly Ile Ala Tyr Phe Ser Met Val Gly Asn Trp 
        355                 360                 365 

Ala Lys Val Leu Val Val Leu Leu Leu Phe Ala Gly Val Asp Ala Glu 
    370                 375                 380 

Thr His Val Thr Gly Gly Ser Ala Gly His Thr Val Ser Gly Phe Val 
385                 390                 395                 400 

Ser Leu Leu Ala Pro Gly Ala Lys Gln Asn Val Gln Leu Ile Asn Thr 
                405                 410                 415 

Asn Gly Ser Trp His Leu Asn Ser Thr Ala Leu Asn Cys Asn Asp Ser 
            420                 425                 430 

Leu Asn Thr Gly Trp Leu Ala Gly Leu Phe Tyr His His Lys Phe Asn 
        435                 440                 445 

Ser Ser Gly Cys Pro Glu Arg Leu Ala Ser Cys Arg Pro Leu Thr Asp 
    450                 455                 460 

Phe Asp Gln Gly Trp Gly Pro Ile Ser Tyr Ala Asn Gly Ser Gly Pro 
465                 470                 475                 480 

Asp Gln Arg Pro Tyr Cys Trp His Tyr Pro Pro Lys Pro Cys Gly Ile 
                485                 490                 495 

Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr Cys Phe Thr Pro Ser 
            500                 505                 510 

Pro Val Val Val Gly Thr Thr Asp Arg Ser Gly Ala Pro Thr Tyr Ser 
        515                 520                 525 

Trp Gly Glu Asn Asp Thr Asp Val Phe Val Leu Asn Asn Thr Arg Pro 
    530                 535                 540 

Pro Leu Gly Asn Trp Phe Gly Cys Thr Trp Met Asn Ser Thr Gly Phe 
545                 550                 555                 560 

Thr Lys Val Cys Gly Ala Pro Pro Cys Val Ile Gly Gly Ala Gly Asn 
                565                 570                 575 

Asn Thr Leu His Cys Pro Thr Asp Cys Phe Arg Lys His Pro Asp Ala 
            580                 585                 590 

Thr Tyr Ser Arg Cys Gly Ser Gly Pro Trp Ile Thr Pro Arg Cys Leu 
        595                 600                 605 

Val Asp Tyr Pro Tyr Arg Leu Trp His Tyr Pro Cys Thr Ile Asn Tyr 
    610                 615                 620 

Thr Ile Phe Lys Ile Arg Met Tyr Val Gly Gly Val Glu His Arg Leu 
625                 630                 635                 640 

Glu Ala Ala Cys Asn Trp Thr Arg Gly Glu Arg Cys Asp Leu Glu Asp 
                645                 650                 655 

Arg Asp Arg Ser Glu Leu Ser Pro Leu Leu Leu Thr Thr Thr Gln Trp 
            660                 665                 670 

Gln Val Leu Pro Cys Ser Phe Thr Thr Leu Pro Ala Leu Ser Thr Gly 
        675                 680                 685 

Leu Ile His Leu His Gln Asn Ile Val Asp Val Gln Tyr Leu Tyr Gly 
    690                 695                 700 

Val Gly Ser Ser Ile Ala Ser Trp Ala Ile Lys Trp Glu Tyr Val Val 
705                 710                 715                 720 

Leu Leu Phe Leu Leu Leu Ala Asp Ala Arg Val Cys Ser Cys Leu Trp 
                725                 730                 735 

Met Met Leu Leu Ile Ser Gln Ala Glu Ala Ala Leu Glu Asn Leu Val 
            740                 745                 750 

Ile Leu Asn Ala Ala Ser Leu Ala Gly Thr His Gly Leu Val Ser Phe 
        755                 760                 765 

Leu Val Phe Phe Cys Phe Ala Trp Tyr Leu Lys Gly Lys Trp Val Pro 
    770                 775                 780 

Gly Ala Val Tyr Thr Phe Tyr Gly Met Trp Pro Leu Leu Leu Leu Leu 
785                 790                 795                 800 

Leu Ala Leu Pro Gln Arg Ala Tyr Ala Leu Asp Thr Glu Val Ala Ala 
                805                 810                 815 

Ser Cys Gly Gly Val Val Leu Val Gly Leu Met Ala Leu Thr Leu Ser 
            820                 825                 830 

Pro Tyr Tyr Lys Arg Tyr Ile Ser Trp Cys Leu Trp Trp Leu Gln Tyr 
        835                 840                 845 

Phe Leu Thr Arg Val Glu Ala Gln Leu His Val Trp Ile Pro Pro Leu 
    850                 855                 860 

Asn Val Arg Gly Gly Arg Asp Ala Val Ile Leu Leu Met Cys Ala Val 
865                 870                 875                 880 

His Pro Thr Leu Val Phe Asp Ile Thr Lys Leu Leu Leu Ala Val Phe 
                885                 890                 895 

Gly Pro Leu Trp Ile Leu Gln Ala Ser Leu Leu Lys Val Pro Tyr Phe 
            900                 905                 910 

Val Arg Val Gln Gly Leu Leu Arg Phe Cys Ala Leu Ala Arg Lys Met 
        915                 920                 925 

Ile Gly Gly His Tyr Val Gln Met Val Ile Ile Lys Leu Gly Ala Leu 
    930                 935                 940 

Thr Gly Thr Tyr Val Tyr Asn His Leu Thr Pro Leu Arg Asp Trp Ala 
945                 950                 955                 960 

His Asn Gly Leu Arg Asp Leu Ala Val Ala Val Glu Pro Val Val Phe 
                965                 970                 975 

Ser Gln Met Glu Thr Lys Leu Ile Thr Trp Gly Ala Asp Thr Ala Ala 
            980                 985                 990 

Cys Gly Asp Ile Ile Asn Gly Leu Pro Val Ser Ala Arg Arg Gly Arg 
        995                 1000                1005 

Glu Ile Leu Leu Gly Pro Ala Asp Gly Met Val Ser Lys Gly Trp Arg 
    1010                1015                1020 

Leu Leu Ala Pro Ile Thr Ala Tyr Ala Gln Gln Thr Arg Gly Leu Leu 
1025                1030                1035                1040 

Gly Cys Ile Ile Thr Ser Leu Thr Gly Arg Asp Lys Asn Gln Val Glu 
                1045                1050                1055 

Gly Glu Val Gln Ile Val Ser Thr Ala Ala Gln Thr Phe Leu Ala Thr 
            1060                1065                1070 

Cys Ile Asn Gly Val Cys Trp Thr Val Tyr His Gly Ala Gly Thr Arg 
        1075                1080                1085 

Thr Ile Ala Ser Pro Lys Gly Pro Val Ile Gln Met Tyr Thr Asn Val 
    1090                1095                1100 

Asp Gln Asp Leu Val Gly Trp Pro Ala Pro Gln Gly Ser Arg Ser Leu 
1105                1110                1115                1120 

Thr Pro Cys Thr Cys Gly Ser Ser Asp Leu Tyr Leu Val Thr Arg His 
                1125                1130                1135 

Ala Asp Val Ile Pro Val Arg Arg Arg Gly Asp Ser Arg Gly Ser Leu 
            1140                1145                1150 

Leu Ser Pro Arg Pro Ile Ser Tyr Leu Lys Gly Ser Ser Gly Gly Pro 
        1155                1160                1165 

Leu Leu Cys Pro Ala Gly His Ala Val Gly Ile Phe Arg Ala Ala Val 
    1170                1175                1180 

Cys Thr Arg Gly Val Ala Lys Ala Val Asp Phe Ile Pro Val Glu Asn 
1185                1190                1195                1200 

Leu Glu Thr Thr Met Arg Ser Pro Val Phe Thr Asp Asn Ser Ser Pro 
                1205                1210                1215 

Pro Val Val Pro Gln Ser Phe Gln Val Ala His Leu His Ala Pro Thr 
            1220                1225                1230 

Gly Ser Gly Lys Ser Thr Lys Val Pro Ala Ala Tyr Ala Ala Gln Gly 
        1235                1240                1245 

Tyr Lys Val Leu Val Leu Asn Pro Ser Val Ala Ala Thr Leu Gly Phe 
    1250                1255                1260 

Gly Ala Tyr Met Ser Lys Ala His Gly Ile Asp Pro Asn Ile Arg Thr 
1265                1270                1275                1280 

Gly Val Arg Thr Ile Thr Thr Gly Ser Pro Ile Thr Tyr Ser Thr Tyr 
                1285                1290                1295 

Gly Lys Phe Leu Ala Asp Gly Gly Cys Ser Gly Gly Ala Tyr Asp Ile 
            1300                1305                1310 

Ile Ile Cys Asp Glu Cys His Ser Thr Asp Ala Thr Ser Ile Leu Gly 
        1315                1320                1325 

Ile Gly Thr Val Leu Asp Gln Ala Glu Thr Ala Gly Ala Arg Leu Val 
    1330                1335                1340 

Val Leu Ala Thr Ala Thr Pro Pro Gly Ser Val Thr Val Pro His Pro 
1345                1350                1355                1360 

Asn Ile Glu Glu Val Ala Leu Ser Thr Thr Gly Glu Ile Pro Phe Tyr 
                1365                1370                1375 

Gly Lys Ala Ile Pro Leu Glu Val Ile Lys Gly Gly Arg His Leu Ile 
            1380                1385                1390 

Phe Cys His Ser Lys Lys Lys Cys Asp Glu Leu Ala Ala Lys Leu Val 
        1395                1400                1405 

Ala Leu Gly Ile Asn Ala Val Ala Tyr Tyr Arg Gly Leu Asp Val Ser 
    1410                1415                1420 

Val Ile Pro Thr Ser Gly Asp Val Val Val Val Ala Thr Asp Ala Leu 
1425                1430                1435                1440 

Met Thr Gly Tyr Thr Gly Asp Phe Asp Ser Val Ile Asp Cys Asn Thr 
                1445                1450                1455 

Cys Val Thr Gln Thr Val Asp Phe Ser Leu Asp Pro Thr Phe Thr Ile 
            1460                1465                1470 

Glu Thr Ile Thr Leu Pro Gln Asp Ala Val Ser Arg Thr Gln Arg Arg 
        1475                1480                1485 

Gly Arg Thr Gly Arg Gly Lys Pro Gly Ile Tyr Arg Phe Val Ala Pro 
    1490                1495                1500 

Gly Glu Arg Pro Ser Gly Met Phe Asp Ser Ser Val Leu Cys Glu Cys 
1505                1510                1515                1520 

Tyr Asp Ala Gly Cys Ala Trp Tyr Glu Leu Thr Pro Ala Glu Thr Thr 
                1525                1530                1535 

Val Arg Leu Arg Ala Tyr Met Asn Thr Pro Gly Leu Pro Val Cys Gln 
            1540                1545                1550 

Asp His Leu Glu Phe Trp Glu Gly Val Phe Thr Gly Leu Thr His Ile 
        1555                1560                1565 

Asp Ala His Phe Leu Ser Gln Thr Lys Gln Ser Gly Glu Asn Leu Pro 
    1570                1575                1580 

Tyr Leu Val Ala Tyr Gln Ala Thr Val Cys Ala Arg Ala Gln Ala Pro 
1585                1590                1595                1600 

Pro Pro Ser Trp Asp Gln Met Trp Lys Cys Leu Ile Arg Leu Lys Pro 
                1605                1610                1615 

Thr Leu His Gly Pro Thr Pro Leu Leu Tyr Arg Leu Gly Ala Val Gln 
            1620                1625                1630 

Asn Glu Ile Thr Leu Thr His Pro Val Thr Lys Tyr Ile Met Thr Cys 
        1635                1640                1645 

Met Ser Ala Asp Leu Glu Val Val Thr Ser Thr Trp Val Leu Val Gly 
    1650                1655                1660 

Gly Val Leu Ala Ala Leu Ala Ala Tyr Cys Leu Ser Thr Gly Cys Val 
1665                1670                1675                1680 

Val Ile Val Gly Arg Val Val Leu Ser Gly Lys Pro Ala Ile Ile Pro 
                1685                1690                1695 

Asp Arg Glu Val Leu Tyr Arg Glu Phe Asp Glu Met Glu Glu Cys Ser 
            1700                1705                1710 

Gln His Leu Pro Tyr Ile Glu Gln Gly Met Met Leu Ala Glu Gln Phe 
        1715                1720                1725 

Lys Gln Lys Ala Leu Gly Leu Leu Gln Thr Ala Ser Arg Gln Ala Glu 
    1730                1735                1740 

Val Ile Ala Pro Ala Val Gln Thr Asn Trp Gln Lys Leu Glu Thr Phe 
1745                1750                1755                1760 

Trp Ala Lys His Met Trp Asn Phe Ile Ser Gly Ile Gln Tyr Leu Ala 
                1765                1770                1775 

Gly Leu Ser Thr Leu Pro Gly Asn Pro Ala Ile Ala Ser Leu Met Ala 
            1780                1785                1790 

Phe Thr Ala Ala Val Thr Ser Pro Leu Thr Thr Ser Gln Thr Leu Leu 
        1795                1800                1805 

Phe Asn Ile Leu Gly Gly Trp Val Ala Ala Gln Leu Ala Ala Pro Gly 
    1810                1815                1820 

Ala Ala Thr Ala Phe Val Gly Ala Gly Leu Ala Gly Ala Ala Ile Gly 
1825                1830                1835                1840 

Ser Val Gly Leu Gly Lys Val Leu Ile Asp Ile Leu Ala Gly Tyr Gly 
                1845                1850                1855 

Ala Gly Val Ala Gly Ala Leu Val Ala Phe Lys Ile Met Ser Gly Glu 
            1860                1865                1870 

Val Pro Ser Thr Glu Asp Leu Val Asn Leu Leu Pro Ala Ile Leu Ser 
        1875                1880                1885 

Pro Gly Ala Leu Val Val Gly Val Val Cys Ala Ala Ile Leu Arg Arg 
    1890                1895                1900 

His Val Gly Pro Gly Glu Gly Ala Val Gln Trp Met Asn Arg Leu Ile 
1905                1910                1915                1920 

Ala Phe Ala Ser Arg Gly Asn His Val Ser Pro Thr His Tyr Val Pro 
                1925                1930                1935 

Glu Ser Asp Ala Ala Ala Arg Val Thr Ala Ile Leu Ser Ser Leu Thr 
            1940                1945                1950 

Val Thr Gln Leu Leu Arg Arg Leu His Gln Trp Ile Ser Ser Glu Cys 
        1955                1960                1965 

Thr Thr Pro Cys Ser Gly Ser Trp Leu Arg Asp Ile Trp Asp Trp Ile 
    1970                1975                1980 

Cys Glu Val Leu Ser Asp Phe Lys Thr Trp Leu Lys Ala Lys Leu Met 
1985                1990                1995                2000 

Pro Gln Leu Pro Gly Ile Pro Phe Val Ser Cys Gln Arg Gly Tyr Lys 
                2005                2010                2015 

Gly Val Trp Arg Val Asp Gly Ile Met His Thr Arg Cys His Cys Gly 
            2020                2025                2030 

Ala Glu Ile Thr Gly His Val Lys Asn Gly Thr Met Arg Ile Val Gly 
        2035                2040                2045 

Pro Arg Thr Cys Arg Asn Met Trp Ser Gly Thr Phe Pro Ile Asn Ala 
    2050                2055                2060 

Tyr Thr Thr Gly Pro Cys Thr Pro Leu Pro Ala Pro Asn Tyr Thr Phe 
2065                2070                2075                2080 

Ala Leu Trp Arg Val Ser Ala Glu Glu Tyr Val Glu Ile Arg Gln Val 
                2085                2090                2095 

Gly Asp Phe His Tyr Val Thr Gly Met Thr Thr Asp Asn Leu Lys Cys 
            2100                2105                2110 

Pro Cys Gln Val Pro Ser Pro Glu Phe Phe Thr Glu Leu Asp Gly Val 
        2115                2120                2125 

Arg Leu His Arg Phe Ala Pro Pro Cys Lys Pro Leu Leu Arg Glu Glu 
    2130                2135                2140 

Val Ser Phe Arg Val Gly Leu His Glu Tyr Pro Val Gly Ser Gln Leu 
2145                2150                2155                2160 

Pro Cys Glu Pro Glu Pro Asp Val Ala Val Leu Thr Ser Met Leu Thr 
                2165                2170                2175 

Asp Pro Ser His Ile Thr Ala Glu Ala Ala Gly Arg Arg Leu Ala Arg 
            2180                2185                2190 

Gly Ser Pro Pro Ser Val Ala Ser Ser Ser Ala Ser Gln Leu Ser Ala 
        2195                2200                2205 

Pro Ser Leu Lys Ala Thr Cys Thr Ala Asn His Asp Ser Pro Asp Ala 
    2210                2215                2220 

Glu Leu Ile Glu Ala Asn Leu Leu Trp Arg Gln Glu Met Gly Gly Asn 
2225                2230                2235                2240 

Ile Thr Arg Val Glu Ser Glu Asn Lys Val Val Ile Leu Asp Ser Phe 
                2245                2250                2255 

Asp Pro Leu Val Ala Glu Glu Asp Glu Arg Glu Ile Ser Val Pro Ala 
            2260                2265                2270 

Glu Ile Leu Arg Lys Ser Arg Arg Phe Ala Gln Ala Leu Pro Val Trp 
        2275                2280                2285 

Ala Arg Pro Asp Tyr Asn Pro Pro Leu Val Glu Thr Trp Lys Lys Pro 
    2290                2295                2300 

Asp Tyr Glu Pro Pro Val Val His Gly Cys Pro Leu Pro Pro Pro Lys 
2305                2310                2315                2320 

Ser Pro Pro Val Pro Pro Pro Arg Lys Lys Arg Thr Val Val Leu Thr 
                2325                2330                2335 

Glu Ser Thr Leu Ser Thr Ala Leu Ala Glu Leu Ala Thr Arg Ser Phe 
            2340                2345                2350 

Gly Ser Ser Ser Thr Ser Gly Ile Thr Gly Asp Asn Thr Thr Thr Ser 
        2355                2360                2365 

Ser Glu Pro Ala Pro Ser Gly Cys Pro Pro Asp Ser Asp Ala Glu Ser 
    2370                2375                2380 

Tyr Ser Ser Met Pro Pro Leu Glu Gly Glu Pro Gly Asp Pro Asp Leu 
2385                2390                2395                2400 

Ser Asp Gly Ser Trp Ser Thr Val Ser Ser Glu Ala Asn Ala Glu Asp 
                2405                2410                2415 

Val Val Cys Cys Ser Met Ser Tyr Ser Trp Thr Gly Ala Leu Val Thr 
            2420                2425                2430 

Pro Cys Ala Ala Glu Glu Gln Lys Leu Pro Ile Asn Ala Leu Ser Asn 
        2435                2440                2445 

Ser Leu Leu Arg His His Asn Leu Val Tyr Ser Thr Thr Ser Arg Ser 
    2450                2455                2460 

Ala Cys Gln Arg Gln Lys Lys Val Thr Phe Asp Arg Leu Gln Val Leu 
2465                2470                2475                2480 

Asp Ser His Tyr Gln Asp Val Leu Lys Glu Val Lys Ala Ala Ala Ser 
                2485                2490                2495 

Lys Val Lys Ala Asn Leu Leu Ser Val Glu Glu Ala Cys Ser Leu Thr 
            2500                2505                2510 

Pro Pro His Ser Ala Lys Ser Lys Phe Gly Tyr Gly Ala Lys Asp Val 
        2515                2520                2525 

Arg Cys His Ala Arg Lys Ala Val Thr His Ile Asn Ser Val Trp Lys 
    2530                2535                2540 

Asp Leu Leu Glu Asp Asn Val Thr Pro Ile Asp Thr Thr Ile Met Ala 
2545                2550                2555                2560 

Lys Asn Glu Val Phe Cys Val Gln Pro Glu Lys Gly Gly Arg Lys Pro 
                2565                2570                2575 

Ala Arg Leu Ile Val Phe Pro Asp Leu Gly Val Arg Val Cys Glu Lys 
            2580                2585                2590 

Met Ala Leu Tyr Asp Val Val Thr Lys Leu Pro Leu Ala Val Met Gly 
        2595                2600                2605 

Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly Gln Arg Val Glu Phe Leu 
    2610                2615                2620 

Val Gln Ala Trp Lys Ser Lys Lys Thr Pro Met Gly Phe Ser Tyr Asp 
2625                2630                2635                2640 

Thr Arg Cys Phe Asp Ser Thr Val Thr Glu Ser Asp Ile Arg Thr Glu 
                2645                2650                2655 

Glu Ala Ile Tyr Gln Cys Cys Asp Leu Asp Pro Gln Ala Arg Val Ala 
            2660                2665                2670 

Ile Lys Ser Leu Thr Glu Arg Leu Tyr Val Gly Gly Pro Leu Thr Asn 
        2675                2680                2685 

Ser Arg Gly Glu Asn Cys Gly Tyr Arg Arg Cys Arg Ala Ser Gly Val 
    2690                2695                2700 

Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr Cys Tyr Ile Lys Ala Arg 
2705                2710                2715                2720 

Ala Ala Cys Arg Ala Ala Gly Leu Gln Asp Cys Thr Met Leu Val Cys 
                2725                2730                2735 

Gly Asp Asp Leu Val Val Ile Cys Glu Ser Ala Gly Val Gln Glu Asp 
            2740                2745                2750 

Ala Ala Ser Leu Arg Ala Phe Thr Glu Ala Met Thr Arg Tyr Ser Ala 
        2755                2760                2765 

Pro Pro Gly Asp Pro Pro Gln Pro Glu Tyr Asp Leu Glu Leu Ile Thr 
    2770                2775                2780 

Ser Cys Ser Ser Asn Val Ser Val Ala His Asp Gly Ala Gly Lys Arg 
2785                2790                2795                2800 

Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr Pro Leu Ala Arg Ala Ala 
                2805                2810                2815 

Trp Glu Thr Ala Arg His Thr Pro Val Asn Ser Trp Leu Gly Asn Ile 
            2820                2825                2830 

Ile Met Phe Ala Pro Thr Leu Trp Ala Arg Met Ile Leu Met Thr His 
        2835                2840                2845 

Phe Phe Ser Val Leu Ile Ala Arg Asp Gln Leu Glu Gln Ala Leu Asp 
    2850                2855                2860 

Cys Glu Ile Tyr Gly Ala Cys Tyr Ser Ile Glu Pro Leu Asp Leu Pro 
2865                2870                2875                2880 

Pro Ile Ile Gln Arg Leu His Gly Leu Ser Ala Phe Ser Leu His Ser 
                2885                2890                2895 

Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala Ala Cys Leu Arg Lys Leu 
            2900                2905                2910 

Gly Val Pro Pro Leu Arg Ala Trp Arg His Arg Ala Arg Ser Val Arg 
        2915                2920                2925 

Ala Arg Leu Leu Ala Arg Gly Gly Arg Ala Ala Ile Cys Gly Lys Tyr 
    2930                2935                2940 

Leu Phe Asn Trp Ala Val Arg Thr Lys Leu Lys Leu Thr Pro Ile Ala 
2945                2950                2955                2960 

Ala Ala Gly Gln Leu Asp Leu Ser Gly Trp Phe Thr Ala Gly Tyr Ser 
                2965                2970                2975 

Gly Gly Asp Ile Tyr His Ser Val Ser His Ala Arg Pro Arg Trp Ile 
            2980                2985                2990 

Trp Phe Cys Leu Leu Leu Leu Ala Ala Gly Val Gly Ile Tyr Leu Leu 
        2995                3000                3005 

Pro Asn Arg 
    3010