Abstract:
The present invention relates to a transgenic  C. elegans  which expresses an amyloid precursor protein (APP) or a part thereof, to the transgene itself, to the protein encoded by the transgene, and also to a process for preparing the transgenic  C. elegans  and to its use.

Description:
BACKGROUND OF THE INVENTION 
     1. Field of the Invention 
     The present invention relates to a transgenic  C. elegans  which expresses an amyloid precursor protein (APP) or a part thereof, to the transgene itself, to the protein encoded by the transgene, and also to a process for preparing the transgenic  C. elegans  and to its use. 
     2. Description of Related Art 
     Several publications are referenced in the application. These references describe the state of the art to which this invention pertains, and are incorporated herein by reference. 
     Alzheimer&#39;s disease (morbus Alzheimer) is a neurodegenerative disorder of the brain which, at the cellular level, is accompanied by a massive loss of neurons in the limbic system and in the cerebral cortex. At the molecular level, it is possible to detect protein depositions, so-called plaques, in the affected areas of the brain, which depositions constitute an important feature of Alzheimer&#39;s disease. The protein which most frequently occurs in these plaques is a peptide of from 40 to 42 amino acids in size which is termed the Aβ peptide. This peptide is a cleavage product of a substantially larger protein of from 695 to 751 amino acids, the so-called amyloid precursor protein (APP). 
     APP is an integral transmembrane protein which traverses the lipid double layer once. By far the largest part of the protein is located extracellularly, while the shorter C-terminal domain is directed into the cytosol (FIG.  1 ). The Aβ peptide is shown in dark gray in FIG.  1 . About two thirds of the Aβ peptide are derived from the extracellular domain of APP and about one third from the transmembrane domain. 
     In addition to the APP which is located in the membrane, it is also possible to detect a secreted form of the amyloid precursor protein, which form comprises the large ectodomain of the APP and is termed APPsec (“secreted APP”). APPsec is formed from APP by proteolytic cleavage which is effected by α-secretase. The proteolytic cleavage takes place at a site in the amino acid sequence of APP which lies within the amino acid sequence of the Aβ peptide (after amino acid residue 16 of the Aβ peptide). Proteolysis of APP by the α-secretase consequently rules out the possibility of the Aβ peptide being formed. 
     The Aβ peptide can consequently only be formed from APP by an alternative processing route. It is postulated that two further proteases are involved in this processing route, with one of the proteases, which is termed β-secretase, cutting the APP at the N terminus of the Aβ peptide and the second protease, which is termed γ-secretase, releasing the C terminus of the Aβ peptide (Kang, J. et al., Nature, 325, 733) (FIG.  1 ). 
     It has not as yet been possible to identify any of the three secretases or proteases (α-secretase, β-secretase and γ-secretase). However, knowledge of the secretases is of great interest, in particular within the context of investigations with regard to Alzheimer&#39;s disease and with regard to identifying the proteins involved, which proteins can then in turn be employed as targets in follow-up studies since, on the one hand, inhibition of the β-secretase, and in particular of the γ-secretase, could lead to a decrease in Aβ production and, on the other hand, activation of the α-secretase would increase the processing of APP into APPsec and thereby simultaneously reduce formation of the Aβ peptide. 
     There is a large amount of evidence that the Aβ peptide is a crucial factor in the development of Alzheimer&#39;s disease. Inter alia, Aβ fibrils are postulated to be neurotoxic in cell culture (Yankner, B. A. et al., (1990) Proc Natl Acad Sci USA,87, 9020). Furthermore, the neuropathology which is characteristic of Alzheimer&#39;s disease already appears at the age of 30 in Down&#39;s syndrome patients, who have an additional copy of APP. In this case, it is assumed that overexpression of APP is followed by an increased conversion into the Aβ peptide (Rumble, B. et al., (1989), N. Engl. J. Med., 320,1446). 
     The familial forms of Alzheimer&#39;s disease constitute what is probably the most powerful evidence of the central role of the Aβ peptide. In these forms, there are mutations in the APP gene around the region of the β-secretase and γ-secretase cleavage sites or in two further AD-associated genes (presenilins) which, in cell culture, lead to a substantial increase in Aβ production (Scheuner, D. et al., (1996), Nature Medicine, 2, 864). 
     While  C. elegans  has already been used as a model organism in Alzheimer&#39;s disease, these studies do not relate to the processing of APP into the Aβ peptide. Some of the studies are concerned with two other Alzheimer-associated proteins, i.e. the presenilins. The presenilins are transmembrane proteins which traverse the membrane 6-8 times. They are of great importance in familial cases of Alzheimers since specific mutations in the presenilin genes lead to Alzheimer&#39;s disease. In this connection, it was shown that homologs to the human presenilins (sel-12, spe-4 and hop-1) are present in  C. elegans , with the function of the presenilins being conserved in humans and worm (Levitan D, Greenwald I (1995) Nature 377, 351; Levitan et al.(1996) Proc Natl Acad Sci USA, 93, 14940; Baumeister R (1997) Genes &amp; Function 1, 149; Xiajun Li and Iva Greenwald (1997) Proc Natl Acad Sci USA, 94, 12204). 
     Other studies deal with the APP homolog in  C. elegans , which is termed Apl-1, and with expression of the Aβ peptide in  C. elegans . However, Apl-1 does not possess any region which is homologous with the amino acid sequence of the Aβ peptide;  C. elegans  does not therefore possess any endogenous Aβ peptide (Daigle I, Li C (1993) Proc Natl Acad Sci USA, 90 (24), 12045). 
     C. D. Link, Proc Natl Acad Sci USA (1995) 92, 9368 described the expression of Aβ peptide (but not that of an Aβ precursor protein) in  C. elegans . These studies involve preparing transgenic worms which express an Aβ1-42 peptide (i.e. the Aβ peptide which consists of 42 amino acids) as a fusion protein together with a synthetic signal peptide and under the control of the muscle-specific promoter unc 54. Muscle-specific protein depositions which reacted with anti-β-amyloid antibodies were detected in the studies. 
     Other studies (e.g. C. Link et al. personal communication) relate to investigations of the aggregation and toxicity of the Aβ peptide in the  C. elegans  model system. 
     Transgenic  C. elegans  lines were established in the present study in order to investigate the existence of a processing machinery in  C. elegans  which is involved in the formation of Aβ peptide and to identify potential secretases in this worm. 
     SUMMARY OF THE INVENTION 
     In this invention, APP genes have been transferred into  C. elegans  to create a transgenic  C. elegans  organism. This transgenic  C. elegans  can then be used to investigate the processing machinery involved in the formation of the Aβ peptide and to identify potential secretases. 
     The present invention relates to a transgene (a gene that has been transferred from one species to another by genetic engineering) which contains 
     a) a nucleotide sequence encoding an amyloid precursor protein (APP) or a part thereof, wherein the nucleotide sequence comprising the APP peptide or part thereof, contains, as part of the sequence, a nucleotide sequence comprising a complete Aβ peptide or a part of the Aβ peptide, and 
     b) where appropriate, one or more further coding and/or non-coding nucleotide sequences, and 
     c) a promoter for expression in a cell of the nematode  Caenorhabditis elegans  ( C. elegans ). 
     The nucleotide sequence preferably encodes the 100 carboxyterminal amino acids of APP, beginning with the sequence of the Aβ peptide and ending with the carboxyterminal amino acid of APP (C100 fragment). The APP is preferably one of the isoforms APP695 (695 amino acids), APP751 (751 amino acids), APP770 (770 amino acids) and L-APP. All the isoforms are formed from the same APP gene by means of alternative splicing. In APP695, exons 7 and 8 were removed by splicing, whereas only exon 8 is lacking in APP751 and exon 7 and 8 are present in APP770. In addition to this, other splicing forms of APP exist in which exon 15 has been removed by splicing. These forms are termed L-APP and are likewise present in the forms which are spliced with regard to exons 7 and 8. 
     In one particular embodiment of the invention, the transgene contains the nucleotide sequence SEQ ID NO.: 1 or a part thereof or a sequence homologous to SEQ ID No. 1. 
     The transgene can preferably contain an additional coding nucleotide sequence which is located at the 5′ end of the nucleotide sequence encoding APP or a part thereof. In one particular embodiment of the invention, the additional nucleotide sequence encodes a signal peptide or a part thereof, for example encodes the APP signal peptide (SP) having the amino acid sequence SEQ ID NO.:9 or a part thereof. The sequence from the N terminus of the Aβ peptide to the C terminus of APP consists of 99 amino acids. The APP signal peptide consists of 17 amino acids. When a fusion product comprising the N terminus of the Aβ peptide to the C terminus of APP and the APP signal peptide is cloned, one or more spacer amino acids is/are preferably inserted between these two parts of the fusion product, with preference being given to inserting one amino acid, for example leucine. The C-terminal fragment is therefore given different designations, e.g. C100 (C=C terminus), LC99 (L=leucine), LC1-99, C99 or SPA4CT (SP=signal peptide, A4=Aβ peptide and CT=C terminus). 
     In one particular embodiment of the invention, the transgene contains the nucleotide sequence SEQ ID NO.: 2 or a part thereof and/or the nucleotide sequence SEQ ID NO.: 3 or a part thereof. 
     In addition to this, the transgene can also contain one or more additional non-coding and/or one or more additional coding nucleotide sequences. 
     For example, the transgene can contain, as an additional non-coding nucleotide sequence, a sequence from an intron of the APP gene, e.g. a sequence which is derived from the 42 bp intron of the APP gene and exhibits the sequence SEQ ID NO.: 4. A transgene which contains the nucleotide sequence SEQ ID NO.: 5 is part of the subject-matter of the invention. 
     The transgene also preferably contains one or more gene-regulating sequences for regulating expression of the encoded protein, preferably a constitutive promoter or a promoter which can be regulated. For example, the promoter can be active in the neuronal, muscular or dermal tissue of  C. elegans  or be ubiquitously active in  C. elegans . A promoter can, for example, be selected from the group of the  C. elegans  promoters unc-54, hsp 16-2, unc-119, goa-1 and sel-12. In one particular embodiment of the invention, the transgene contains a promoter having the nucleotide sequence SEQ ID NO.: 6. In one particular embodiment, the transgene contains the nucleotide sequence SEQ ID NO.: 7. 
     The transgene can be present in a vector, for example in an expression vector. For example, a recombinant expression vector can contain the nucleotide sequence SEQ ID NO.: 8. 
     The invention also relates to the preparation of an expression vector, with a transgene being integrated into a vector in accordance with known methods. In particular, the invention relates to the use of an expression vector for preparing a transgenic cell, with it being possible for this cell to be part of a non-human organism, e.g.  C. elegans.    
     The invention also relates to the preparation of the transgene, with suitable part sequences being ligated in the appropriate order and in the correct reading frame, where appropriate while inserting linkers. In particular, the invention relates to the use of the transgene, for example for preparing a transgenic cell, with it being possible for this cell to be part of a non-human organism. For example, the cell can be a  C. elegans  cell. 
     One particular embodiment of the invention relates to a transgenic  C. elegans  which contains the transgene. The transgene can also be present in the  C. elegans  in an expression vector. The transgene can be present in the  C. elegans  intrachromosomally and/or extrachromosomally. One or more transgenes or expression vectors which contain the transgene can be present intrachromosomally and/or extrachromosomally as long tandem arrays. A transgenic cell or a transgenic organism preferably contains another expression vector as well, which vector contains a nucleotide sequence which encodes a marker, with the marker either being a temperature-sensitive marker or a phenotypic marker. For example, the marker can be a visual marker or a behaviorally phenotypic marker. Examples are fluorescent markers, e.g. GFP (green fluorescent protein) or EGFP (enhanced green fluorescent protein), marker genes which encode a dominant, mutated form of a particular protein, e.g. a dominant Rol6 mutation, or marker sequences which encode antisense RNA, e.g. the antisense RNA of Unc-22. 
     One or more copies of the transgene and/or of the expression vector and, where appropriate, of an additional expression vector are preferably present in the germ cells and/or the somatic cells of the transgenic  C. elegans.    
     The invention also relates to a process for preparing a transgenic  C. elegans , with a transgene and/or an expression vector, where appropriate in the presence of an additional expression vector which contains a nucleotide sequence which encodes a marker, being microinjected into the germ cells of a  C. elegans . A DNA construct which expresses SP-C100 (SP=signal peptide) under the control of a neuron-specific promoter can, for example, be used for preparing the transgenic  C. elegans  lines (FIG.  2 ). Since C100 is composed of the Aβ sequence and the C terminus of APP, only the γ-secretase cleavage is required in order to release the Aβ peptide from C100. C100 is also a substrate for the γ-secretase. 
     The invention also relates to the use of a transgenic  C. elegans , for example for expressing an SP-C100 fusion protein. An SP-C100 fusion protein having the amino acid sequence SEQ ID NO.: 10 is part of the subject-matter of the invention. 
     In particular, the invention relates to the use of a transgenic  C. elegans  for identifying a γ-secretase activity and/or an α-secretase activity in  C. elegans , to its use in methods for identifying and/or characterizing substances which inhibit the γ-secretase activity, to its use in methods for identifying and/or characterizing substances which increase the α-secretase activity, and to its use in methods for identifying and/or characterizing substances which can be used as active compounds for treating and/or preventing Alzheimer&#39;s disease. 
     DESCRIPTION OF THE PREFERRED EMBODIMENTS 
     In the present study, the nematode  Caenorhabditis elegans  ( C. elegans ) was chosen as the model organism for identifying secretases which are involved in processing APP into the Aβ peptide. This worm is outstandingly suitable for genetic studies and has therefore in the past been employed on many occasions for investigating universally important processes such as programmed cell death, neuronal guidance and RAS/MAP kinase signaling (Riddle, D. L. et al. (1997)). 
     The important points which make  C. elegans  especially appropriate for such studies include the following (C. Kenyon, Science (1988) 240, 1448; P. E. Kuwabara (1997), TIG, 13, 454): 
     Its small genome, which is composed of about 19,000 genes or 97 Mb and which was sequenced completely in December 1998. (The  C. elegans  Sequencing Consortium, Science (1998), 282, 2012). 
     Its reproduction by self fertilization. In the case of the two sexes of  C. elegans , a distinction is made between males and hermaphrodites, i.e. hermaphroditic animals which fertilize their eggs themselves before laying. A crucial advantage of this type of reproduction is that, after a transgene has been introduced into the germ line, a hermaphrodite can automatically generate homozygous transgenic descendants. There is therefore no need for any further crossing steps, as in the case of Drosophila, for example, for preparing transgenic lines. 
     Its easy handling in the laboratory due to its small size (about 1 mm in length) and its relatively undemanding growth conditions. As a result, a large number of worms can be handled routinely in the laboratory. 
     Its short generation time of 3 days, which makes it possible to obtain large quantities of biological material for analysis within a very short time. 
     A complete cell description for the development and anatomy of  C. elegans  is available. 
     Detailed genetic maps and methods for genetic analysis in  C. elegans  are available. 
     Technologies for preparing knock-out animals are available. In the same way, technologies exist for mutagenizing the  C. elegans  genome (transposon mutagenesis and ethyl methanesulfonate (EMS) mutagenesis). 
     The following are possible uses of the transgenic  C. elegans  lines: 
     1. Identification of a γ-secretase-like activity in  C. elegans  using mutagenesis approaches. It is planned that a transposon mutagenesis, which destroys the γ-secretase-like activity, should be carried out and that the corresponding gene should be sought by detecting the worms which no longer possess this activity. Such a screening method is described in the literature: Korswagen H. C. et al., (1996), 93, 14680 Proc Natl Acad Sci USA. 
     Alternative approaches would be mutagenesis using ethyl methanesulfonate (EMS) or else anti-sense RNA approaches. In the latter case, an attempt could be made to find motifs which were common to all  C. elegans  proteases and to downregulate these proteases specifically using anti-sense RNAs which were directed against these motifs. Screening for the Aβ peptide could then show whether one of the proteases was involved in Aβ peptide production. 
     2. Identification of a γ-secretase-like activity in  C. elegans , perhaps by a similar route to that described in item 1. 
     3. Armed with knowledge of a γ-secretase or γ-secretase-like activity in  C. elegans , it is possible to search for human γ-secretase or γ-secretase-like activity by means of a homology comparison. 
     4. Identification of drugs which 
     inhibit the activity of γ-secretase, in order to inhibit Aβ production from the amyloid precursor protein directly. 
     activate γ-secretase and thereby indirectly inhibit formation of the Aβ peptide by increasing APPsec production. 
     This approach could take place in a 96-well format since  C. elegans  can be maintained in suspension in 96-well plates. 
     Since the screening is carried out on a whole organism, it is possible, to a large extent, to exclude drugs which have an unspecific toxic effect. 
     5. Investigation of the aggregation behavior, and of a possible neurotoxic effect, of the Aβ peptide in  C. elegans . Screening for drugs which inhibit aggregation of the Aβ peptide. 
     6. Investigation of the modulation of APP processing by other proteins (e.g. presenilins or ApoE) as a result of their overexpression or knock-out. Since the presenilins are Alzheimer-associated proteins and ApoE constitutes a risk factor in Alzheimer&#39;s disease, these proteins could have an effect on formation of the Aβ peptide and, as a consequence, their role in the APP processing pathway could be investigated. 
     7. Where appropriate, validation of an α-secretase and/or γ-secretase activity which has been found using other experimental approaches known to the skilled person. 
    
    
     FIG.  1 : FIG. 1 shows the amyloid precursor protein (APP695 isoform and APP770 and APP751 isoforms) and secretase cleavage products. 
     FIG.  2 : FIG. 2 describes the construction of the transgenic vector “Unc-119-SP-C100”, which contains an unc-119 promoter, an APP signal peptide and the C100 fragment from APP, with “unc-119” being a neuron-specific  C. elegans  promoter, the APP signal peptide corresponding to amino acids 1 to 24 of APP and C100 corresponding to the 100 C-terminal amino acids of APP (=C100). C100 is composed of the Aβ sequence and the C terminus of APP (Shoji, M et al., (1992) Science 258, 126). The vector Unc-119-SP-C100 possesses 5112 base pairs. 
    
    
     EXAMPLES 
     The following examples are illustrative of some of the products and compositions and methods of making and using the same falling within the scope of the present invention. 
     Example 1 
     Preparing an Expression Vector Which Contains the Transgene 
     Two vectors, i.e. pSKLC1-99, which encodes SP-C100, and pBY103, which contains the unc-119 promoter, were used for the cloning, with the SP-C100-encoding DNA being cloned into the pBY103 vector behind the unc-119 promoter. The basic vector pBY103 is composed of the vector backbone pPD49.26, which is described in “ Caenorhabditis elegans : Modern Biological Analysis of an Organism” (1995) Ed. Epstein et al., Vol 48, pp. 473, into which the unc-119 promoter (Maduro et al. Genetics (1995), 141, p. 977) has been cloned by way of the HindIII/BamHI sites. The plasmid unc-119-SP-C100 was prepared by KpnI/SacI digestion of pSKLC1-99 and cloning of the LC99 fragment into pBY103 (Shoji et al. (1992). 
     Example 2 
     Preparing the Transgenic  C. elegans  Lines 
     The method of microinjection was used for preparing the transgenic  C. elegans  lines (Mello et al., (1991) EMBO J. 10 (12) 3959; C. Mello and A. Fire, Methods in Cell Biology, Academic Press Vol. 48, pp. 451, 1995; C. D. Link, Proc Natl Acad Sci USA (1995) 92, 9368). 
     Two different  C. elegans  strains, i.e. wild-type N2 and him-8 (high incidence of males), were used. The unc-119-SP-C100 construct was microinjected into the gonads of young adult hermaphrodites using a microinjection appliance. The DNA concentration was about 20 ng/μl. 
     A marker plasmid was injected together with the unc-119-SP-C100 construct. This marker plasmid is the plasmid ttx3-GFP, which encodes the green fluorescent protein under the control of the ttx3 promoter. The activity of the ttx3 promoter is specific for particular neurons of the  C. elegans  head, the so-called AIY neurons, which play a role in the thermotaxis of the worm. 
     When plasmid DNA is microinjected, it is assumed that long tandem arrays, which are composed of many copies of plasmid DNA (in our case, of the ttx3-GFP plasmid and the unc-119-SP-C100 plasmid), are formed by recombination. A certain percentage of these arrays integrate into the  C. elegans  genome. However, the arrays are more likely to be present extrachromosomally. 
     Worms which had been injected successfully exhibit a green fluorescence in the AIY neurons of the head region when stimulated with light of a wavelength of about 480 nm. It was possible to detect such nematodes. 
     Example 3 
     Describing the C100 Transgenic  C. elegans  Lines 
     1. Phenotypic Features 
     Following stimulation with light of a wavelength of 480 nm, C100-transgenic worms exhibit a green fluorescence in the AIY neurons of the head region. Since it was also possible to detect green fluorescence in the head neurons once again in the descendants of the worms, it can be assumed that the plasmids are able to pass down through the germ line. However, the penetrance is not 100%, which makes it possible to conclude that the long tandem arrays composed of ttx3-GFP marker DNA and unc-119-SP-C100 are present extrachromosomally rather than being integrated into the genome. 
     Example 4 
     Detecting C100 Expression in a Blot 
     Six different transgenic C100  C. elegans  lines (three in an N2 wt background and three in a him 8 background) were examined in a Western blot for expression of the C100 fragment using a polyclonal antiserum directed against the C terminus of APP. A band having the appropriate molecular weight of about 10 kDa was detectable in all the six lines. 
     Example 5 
     Detecting the C100 in an ELISA 
     In an Aβ Sandwich ELISA, signals which were above the background level, and which were statistically significant in two cases, were detected in cell extracts from transgenic animals. This indicates that  C. elegans  could possess a γ-secretase-like activity. 
     In the Aβ Sandwich ELISA assay, 96-well plates are first of all incubated with the monoclonal antibody clone 6E10 (SENETEK PLC., MO, USA), which reacts specifically with the Aβ peptide (amino acids 1-17), and then coated with worm extracts from transgenic worms or control worms. The Aβ peptide is detected using the monoclonal Aβ antibody 4G8 (SENETEK PLC., MO, USA), which recognizes amino acids 17-24 in the Aβ peptide and is labeled with biotin. The detection is effected by way of the alkaline phosphatase reaction using an appropriate antibody which is directed against biotin. Disruption of the worms involves detergent treatment, nitrogen shock freezing, sonication and rupture of the cells using glass beads. 
     The ELISA signal from the above-described experiment can be based either on weak expression of the Aβ peptide or on expression of the C100 precursor protein, since the appropriate epitopes are present in both proteins. 
     Expression of the Aβ peptide could, for example, also be specifically detected in an analogous manner: for this, Aβ-specific antibodies which do not react with the C100 precursor would have to be employed in an Aβ Sandwich ELISA. An Aβ-specific antibody could, for example, be a monoclonal antibody which specifically recognizes the C-terminal end of the Aβ form, which is composed of 40 or 42 amino acids. In parallel, the Aβ peptide could be detected in a Western blot using the monoclonal antibodies 4G8 and 6E10 and then be distinguished from the larger C100 precursor by its molecular weight of 4 kD. 
     The vectors can be obtained from Andrew Fire (Department of Embryology, Carnegie Institution of Washington, Baltimore, Md. 21210, USA) in the case of pPD49.26 and LC99 (amyloid precursor protein), which is deposited under ATCC number 106372. The unc-119 promoter can be obtained from Maduro, M. (Department of Biological Science, Universitiy of Alberta Edmonton, Canada), while unc-54 and unc-16.2 can be obtained from Andrew Fire. 
     The above description of the invention is intended to be illustrative and not limiting. Various changes or modifications in the embodiments described may occur to those skilled in the art. These can be made without departing from the spirit or scope of the invention. 
     
       
         
               
               
             
           
               
                 SEQ ID NO.1: Nucleotide sequence of C100 
                   
               
               
                   
               
               
                 CTGGATGC AGAATTCCGA CATGACTCAG GATATGAAGT TCATCATCAAAAATTGGTGT 
               
               
                   
               
               
                 TCTTTGCAGA AGATGTGGGT TCAAACAAAG GTGCAATCAT TGGACTCATGGTGGGCGGTG 
               
               
                   
               
               
                 TTGTCATAGC GACAGTGATC GTCATCACCT TGGTGATGCT GAAGAAGAAACAGTACACAT 
               
               
                   
               
               
                 CCATTCATCA TGGTGTGGTG GAGGTTGACG CCGCTGTCAC CCCAGAGGAGCGCCACCTGT 
               
               
                   
               
               
                 CCAAGATGCA GCAGAACGGC TACGAAAATC CAACCTACAA GTTCTTTGAGCAGATGCAGA ACTAG 
               
               
                   
               
               
                 SEQ ID NO.2: Nucleotide sequence of SP 
               
               
                   
               
               
                 ATG CTGCCCGGTT TGGCACTGTT CCTGCTGGCC GCCTGGACGG CTCGGGCG 
               
               
                   
               
               
                 SEQ ID NO.3: Nucleotide sequence of SP+C100 
               
               
                   
               
               
                 ATG CTGCCCGGTT TGGCACTGTT CCTGCTGGCC GCCTGGACGG CTCGGGCGCT G 
               
               
                   
               
               
                 GATGC AGAATTCCGA CATGACTCAG GATATGAAGT TCATCATCAA AAATTGGTGT 
               
               
                   
               
               
                 TCTTTGCAGA AGATGTGGGT TCAAACAAAG GTGCAATCAT TGGACTCATG 
               
               
                   
               
               
                 GTGGGCGGTG TTGTCATAGC GACAGTGATC GTCATCACCT TGGTGATGCT GAAGAAGAAA 
               
               
                   
               
               
                 CAGTACACAT CCATTCATCA TGGTGTGGTG GAGGTTGACG CCGCTGTCAC CCCAGAGGAG 
               
               
                   
               
               
                 CGCCACCTGT CCAAGATGCA GCAGAACGGC TACGAAAATC CAACCTACAA GTTCTTTGAG 
               
               
                   
               
               
                 CAGATGCAGA ACTAG 
               
               
                   
               
               
                 SEQ ID NO.4: Nucleotide sequence of the 42bp intron 
               
               
                   
               
               
                 GTATGTTTCGAATGATACTAACATAACATAGAACATTTTCAG 
               
               
                   
               
               
                 SEQ ID NO.5: Nucleotide sequence of intron+SP+C100 
               
               
                   
               
               
                 GTATGTTTCGAATGATACTAACATAACATAGAACATTTTCAGGAGGACCCTTGGCTAGCGTCGACGGT 
               
               
                   
               
               
                 ACCGGGCCCCCCCTCGAGGTCGACGGTATCGATAACCTTCACAGCAGCGCACTCGGTGCCCCGCG 
               
               
                   
               
               
                 CAGGGTCGCGATG CTGCCCGGTT TGGCACTGTT CCTGCTGGCCGCCTGGACGG CTCGGGCGCT 
               
               
                   
               
               
                 GGATGC AGAATTCCGAATGACTCAGGATATGAAGTCATCATCAAAAATTGGTGT TCTTTGCAGA 
               
               
                   
               
               
                 AGATGTGGGT TCAAACAAAG GTGCAATCAT TGGACTCATG GTGGGCGGTG TTGTCATAGC 
               
               
                   
               
               
                 GACAGTGATC GTCATCACCT TGGTGATGCT GAAGAAGAAA CAGTACACAT CCATTCATCA 
               
               
                   
               
               
                 TGGTGTGGTG GAGGTTGACG CCGCTGTCAC CCCAGAGGAGCGCCACCTGT CCAAGATGCA 
               
               
                   
               
               
                 GCAGAACGGC TACGAAAATC CAACCTACAA GTTCTTTGAG CAGATGCAGA ACTAG 
               
               
                   
               
               
                 SEQ ID NO.6: Nucleotide sequence of unc-119 
               
               
                   
               
               
                 AAGCTTCAGTAAAAGAAGTAGAATTTTATAGTTTTTTTTCTGTTTGAAAAATTCTCCCCATCAATGTTCT 
               
               
                   
               
               
                 TTCAAATAAATACATCACTAATGCAAAGTATTCTATAACCTCATATCTAAATTCTTCAAAATCTTAACAT 
               
               
                   
               
               
                 ATC 
               
               
                   
               
               
                 TTATCATTGCTTTAAGTCAACGTAACATTAAAAAAAATGTTTTGGAAAATGTGTCAAGTCTCTCAAAATT 
               
               
                   
               
               
                 CAGTTTTTTAAACCACTCCTATAGTCCTATAGTCCTATAGTTACCCATGAAATCCTTATATATTACTGTA 
               
               
                   
               
               
                 AAATGTTTCAAAAACCATTGGCAAATTGCCAGAACTGAAAATTTCCGGCAAATTGGGGAACCGGCAA 
               
               
                   
               
               
                 ATTGCCAATTTGCTGAATTTGCCGGAAACGGTAATTGCCGAAAGTTTTTGACACGAAAATGGCAAATT 
               
               
                   
               
               
                 GTGGTTTTAAAATTTTTTTTTTTGGAAATTTCAGAATTTCAATTTTAATCGGCAAAACTGTAGGCATCCT 
               
               
                   
               
               
                 AAGAATGTTCCTACATCTATTTTGAAAAGTAAGCGAATTAATTCTATGAAAATGTCTAAAGAAAATGGG 
               
               
                   
               
               
                 GAAACAATTTCAAAAAGGCACAGTTTCAATGGTTTCCGAATTATACTAAATCCCTCTAAAAACTTCCGG 
               
               
                   
               
               
                 CAAATTGATATCCGTAAAAGAGCAAATCCGCATTTTTGCCGAAAATTAAAATTTCCGACAAATCGGCA 
               
               
                   
               
               
                 AACCGGCAATTTGGCGAAATTTGCCGGAACGATTGCCGCCCACCCCTGTTCCAGAGGTTCAAACTG 
               
               
                   
               
               
                 GTAGCAAAGCTCAAAATTTCTCAAATTCTCCAATTTTTTTTTGAATTTTGGCAGTGTACCAAAATGACA 
               
               
                   
               
               
                 TTCAGTCATATTGGTTTATTATAGATTTATTTAGATAAAATCCTAAATGATTCTACCTTTAAAGATGCCC 
               
               
                   
               
               
                 ACTTTAAAAGTAATGACTCAAACTTCAAATTGCTCTAAGATTCTATTGAATTACCATCTTTTCCTCTCAT 
               
               
                   
               
               
                 TTTCTCTCACTGTCTATTTCATCACAAATTCATCCCTCTCTCCTCTCTTCTCTCTCCCTCTCTCTCTCTT 
               
               
                   
               
               
                 TCTCTTTGCTCATCATCTGTCATTTTGTCCGTTCCTCTCTCTGCGCCCTCAGCGTTCCCCACACTCTC 
               
               
                   
               
               
                 TCGCTTCTCTTTTCCTAGACGTCTTCTTTTTTCATCTTCTTCAGCCTTTTTCGCCATTTTCCATCTCTGT 
               
               
                   
               
               
                 CAATCATTACGGACGACCCCCATTATCGAT 
               
               
                   
               
               
                 SEQ ID NO.7: Nucleotide sequence of unc-119+intron+SP+C100 
               
               
                   
               
               
                 AAGCTTCAGTAAAAGAAGTAGAATTTTATAGTTTTTTTTCTGTTTGAAAAATTCTCCCCATCA 
               
               
                   
               
               
                 ATGTTCTTTCAAATAAATACATCACTAATGCAAAGTATTCTATAACCTCATATCTAAATTCTTCAAAATC 
               
               
                   
               
               
                 TTAACATATCTTATCATTGCTTTAAGTCAACGTAACATTAAAAAAAATGTTTTGGAAAATGTGTCAAGTC 
               
               
                   
               
               
                 TCTCAAAATTCAGTTTTTTAAACCACTCCTATAGTCCTATAGTCCTATAGTTACCCATGAAATCCTTATA 
               
               
                   
               
               
                 TATTACTGTAAAATGTTTCAAAAACCATTGGCAAATTGCCAGAACTGAAAATTTCCGGCAAATTGGGG 
               
               
                   
               
               
                 AACCGGCAAATTGCCAATTTGCTGAATTTGCCGGAAACGGTAATTGCCGAAAGTTTTTGACACGAAAA 
               
               
                   
               
               
                 TGGCAAATTGTGGTTTTAAAATTTTTTTTTTTGGAAATTTCAGAATTTCAATTTTAATCGGCAAAACTGT 
               
               
                   
               
               
                 AGGCATCCTAAGAATGTTCCTACATCTATTTTGAAAAGTAAGCGAATTAATTCTATGAAAATGTCTAAA 
               
               
                   
               
               
                 GAAAATGGGGAAACAATTTCAAAAAGGCACAGTTTCAATGGTTTCCGAATTATACTAAATCCCTCTAA 
               
               
                   
               
               
                 AAACTTCCGGCAAATTGATATCCGTAAAAGAGCAAATCCGCATTTTTGCCGAAAATTAAAATTTCCGA 
               
               
                   
               
               
                 CAAATCGGCAAACCGGCAATTTGGCGAAATTTGCCGGAACGATTGCCGCCCACCCCTGTTCCAGAG 
               
               
                   
               
               
                 GTTCAAACTGGTAGCAAAGCTCAAAATTTCTCAAATTCTCCAATTTTTTTTTGAATTTTGGCAGTGTAC 
               
               
                   
               
               
                 CAAAATGACATTCAGTCATATTGGTTTATTATAGATTTATTTAGATAAAATCCTAAATGATTCTACCTTT 
               
               
                   
               
               
                 AAAGATGCCCACTTTAAAAGTAATGACTCAAACTTCAAATTGCTCTAAGATTCTATTGAATTACCATCT 
               
               
                   
               
               
                 TTTCCTCTCATTTTCTCTCACTGTCTATTTCATCACAAATTCATCCCTCTCTCCTCTCTTCTCTCTCCCT 
               
               
                   
               
               
                 CTCTCTCTCTTTCTCTTTGCTCATCATCTGTCAT 
               
               
                   
               
               
                 TTTGTCCGTTCCTCTCTCTGCGCCCTCAGCGTTCCCCACACTCTCTCGCTTCTCTTTTCCTAGACGTC 
               
               
                   
               
               
                 TTCTTTTTTCATCTTCTTCAGCCTTTTTCGCCATTTTCCATCTCTGTCAATCATTACGGACGACCCCCA 
               
               
                   
               
               
                 TTATCGATAAGATCTCCACGGTGGCCGCGAATTCCTGCAGCCCGGGGGATCCCCGGGATTGGCCAA 
               
               
                   
               
               
                 AGGACCCAAAGGTATGTTTCGAATGATACTAACATAACATAGAACATTTTCAGGAGGACCCTTGGCTA 
               
               
                   
               
               
                 GCGTCGACGGTACCGGGCCCCCCCTCGAGGTCGACGGTATCGATAACCTTCACAGCAGCGCACTC 
               
               
                   
               
               
                 GGTGCCCCGCGCAGGGTCGCGATGCTGCCCGGTT 
               
               
                   
               
               
                 TGGCACTGTTCCTGCTGGCCGCCTGGACGGCTCGGGCGCTGGATGCAGAATTCCGA 
               
               
                   
               
               
                 CATGACTCAGGATATGAAGTTCATCATCAAAAATTGGTGTTCTTTGCAGAAGATGTGGGTTCAAACAA 
               
               
                   
               
               
                 AG GTGCAATCAT TGGACTCATGGTGGGCGGTGTTGTCATAGCGACAGTGATCGTCATCACCT 
               
               
                   
               
               
                 TGGTGATGCT GAAGAAGAAACAGTACACAT CCATTCATCA TGGTGTGGTG GAGGTTGACG 
               
               
                   
               
               
                 CCGCTGTCAC CCCAGAGGAGCGCCACCTGT CCAAGATGCA GCAGAACGGC TACGAAAATC 
               
               
                   
               
               
                 CAACCTACAA GTTCTTTGAGCAGATGCAGA ACTAG 
               
               
                   
               
               
                 SEQ ID NO.8: Nucleotide sequence of the expression vector 
               
               
                   
               
               
                 ACCCCCGCCACAGCAGCCTCTGAAGTTGGACACGGATCCACTAGTTCTAGAGCGGCCGCCACCGC 
               
               
                   
               
               
                 GGTGGAGCTCCGCATCGGCCGCTGTCATCAGATCGCCATCTCGCGCCCGTGCCTCTGACTTCTAAG 
               
               
                   
               
               
                 TCCAATTACTCTTCAACATCCCTACATGCTCTTTCTCCCTGTGCTCCCACCCCCTATTTTTGTTATTAT 
               
               
                   
               
               
                 CAAAAAAACTTCTTCTTAATTTCTTTGTTTTTTAGCTTCTTTTAAGTCACCTCTAACAATGAAATTGTGT 
               
               
                   
               
               
                 AGATTCAAAAATAGAATTAATTCGTAATAAAAAGTCGAAAAAAATTGTGCTCCCTCCCCCCATTAATAA 
               
               
                   
               
               
                 TAATTCTATCCCAAAATCTACACAATGTTCTGTGTACACTTCTTATGTTTTTTTTACTTCTGATAAATTTT 
               
               
                   
               
               
                 TTTTGAAACATCATAGAAAAAACCGCACACAAAATACCTTATCATATGTTACGTTTCAGTTTATGACCG 
               
               
                   
               
               
                 CAATTTTTATTTCTTCGCACGTCTGGGCCTCTCATGACGTCAAATCATGCTCATCGTGAAAAAGTTTT 
               
               
                   
               
               
                 GGAGTATTTTTGGAATTTTTCAATCAAGTGAAAGTTTATGAAATTAATTTTCCTGCTTTTGCTTTTTGGG 
               
               
                   
               
               
                 GGTTTCCCCTATTGTTTGTCAAGAGTTTCGAGGACGGCGTTTTTCTTGCTAAAATCACAAGTATTGAT 
               
               
                   
               
               
                 GAGCACGATGCAAGAAAGATCGGAAGAAGGTTTGGGTTTGAGGCTCAGTGGAAGGTGAGTAGAAGT 
               
               
                   
               
               
                 TGATAATTTGAAAGTGGAGTAGTGTCTATGGGGTTTTTGCCTTAAATGACAGAATACATTCCCAATATA 
               
               
                   
               
               
                 CCAAACATAACTGTTTCCTACTAGTCGGCCGTACGGGCCCTTTCGTCTCGCGCGTTTCGGTGATGAC 
               
               
                   
               
               
                 GGTGAAAACCTCTGACACATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGG 
               
               
                   
               
               
                 AGCAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCTGGCTTAACTATGC 
               
               
                   
               
               
                 GGCATCAGAGCAGATTGTACTGAGAGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAG 
               
               
                   
               
               
                 GAGAAAATACCGCATCAGGCGGCCTTAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCAT 
               
               
                   
               
               
                 GATAATAATGGTTTCTTAGACGTCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGT 
               
               
                   
               
               
                 TTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAAT 
               
               
                   
               
               
                 ATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTT 
               
               
                   
               
               
                 GCCTTCCTGTTTTTGCTC 
               
               
                   
               
               
                 ACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCG 
               
               
                   
               
               
                 AACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAG 
               
               
                   
               
               
                 CACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGT 
               
               
                   
               
               
                 CGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGG 
               
               
                   
               
               
                 ATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTT 
               
               
                   
               
               
                 ACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTA 
               
               
                   
               
               
                 ACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACG 
               
               
                   
               
               
                 ATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCC 
               
               
                   
               
               
                 GGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTC 
               
               
                   
               
               
                 CGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAG 
               
               
                   
               
               
                 CACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTA 
               
               
                   
               
               
                 TGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGA 
               
               
                   
               
               
                 CCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAG 
               
               
                   
               
               
                 ATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCC 
               
               
                   
               
               
                 CGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAA 
               
               
                   
               
               
                 AAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGT 
               
               
                   
               
               
                 AACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCAC 
               
               
                   
               
               
                 TTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCA 
               
               
                   
               
               
                 GTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGT 
               
               
                   
               
               
                 CGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGA 
               
               
                   
               
               
                 TACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCG 
               
               
                   
               
               
                 GTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCT 
               
               
                   
               
               
                 TTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGG 
               
               
                   
               
               
                 CGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTG 
               
               
                   
               
               
                 CTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCT 
               
               
                   
               
               
                 GATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGC 
               
               
                   
               
               
                 GCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGT 
               
               
                   
               
               
                 TTCCGGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCAC 
               
               
                   
               
               
                 CCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCAC 
               
               
                   
               
               
                 ACAGGAAACAGCTATGACCATGATTACGCCAAGCTT 
               
               
                   
               
               
                 SEQ ID NO.9: Amino acid sequence of SP 
               
               
                   
               
               
                 MLPGLALFLL AAWTARA 
               
               
                   
               
               
                 SEQ ID NO.10: Amino acid sequence of the fusion protein 
               
               
                   
               
               
                 MLPGLALFLL AAWTARALDA EFRHDSGYEV HHQKLVFFAE DVGSNKGAII 
               
               
                   
               
               
                 GLMVGGVVIA TVIVITLVML KKKQYTSIHH GVVEVDAAVT PEERHLSKMQ 
               
               
                   
               
               
                 QNGYENPTYK FFEQMQN 
               
               
                   
               
               
                 SEQ ID NO. 11: Nucleotide sequence of the vector unc-119-SP-C100 
               
               
                   
               
               
                 ATGACCATGATTACGCCAAGCTTCAGTAAAAGAAGTAGAATTTTATAGTTTTTTTTCTGTTTGAAAAAT 
               
               
                   
               
               
                 TCTCCCCATCAATGTTCTTTCAAATAAATACATCACTAATGCAAAGTATTCTATAACCTCATATCTAAAT 
               
               
                   
               
               
                 TCTTCAAAATCTTAACATATCTTATCATTGCTTTAAGTCAACGTAACATTAAAAAAAATGTTTTGGAAAA 
               
               
                   
               
               
                 TGTGTCAAGTCTCTCAAAATTCAGTTTTTTAAACCACTCCTATAGTCCTATAGTCCTATAGTTACCCAT 
               
               
                   
               
               
                 GAAATCCTTATATATTACTGTAAAATGTTTCAAAAACCATTGGCAAATTGCCAGAACTGAAAATTTCCG 
               
               
                   
               
               
                 GCAAATTGGGGAACCGGCAAATTGCCAATTTGCTGAATTTGCCGGAAACGGTAATTGCCGAAAGTTT 
               
               
                   
               
               
                 TTGACACGAAAATGGCAAATTGTGGTTTTAAAATTTTTTTTTTTGGAAATTTCAGAATTTCAATTTTAAT 
               
               
                   
               
               
                 CGGCAAAACTGTAGGCATCCTAAGAATGTTCCTACATCTATTTTGAAAAGTAAGCGAATTAATTCTAT 
               
               
                   
               
               
                 GAAAATGTCTAAAGAAAATGGGGAAACAATTTCAAAAAGGCACAGTTTCAATGGTTTCCGAATTATAC 
               
               
                   
               
               
                 TAAATCCCTCTAAAAACTTCCGGCAAATTGATATCCGTAAAAGAGCAAATCCGCATTTTTGCCGAAAA 
               
               
                   
               
               
                 TTAAAATTTCCGACAAATCGGCAAACCGGCAATTTGGCGAAATTTGCCGGAACGATTGCCGCCCACC 
               
               
                   
               
               
                 CCTGTTCCAGAGGTTCAAACTGGTAGCAAAGCTCAAAATTTCTCAAATTCTCCAATTTTTTTTTGAATT 
               
               
                   
               
               
                 TTGGCAGTGTACCAAAATGACATTCAGTCATATTGGTTTATTATAGATTTATTTAGATAAAATCCTAAAT 
               
               
                   
               
               
                 GATTCTACCTTTAAAGATGCCCACTTTAAAAGTAATGACTCAAACTTCAAATTGCTCTAAGATTCTATT 
               
               
                   
               
               
                 GAATTACCATCTTTTCCTCTCATTTTCTCTCACTGTCTATTTCATCACAAATTCATCCCTCTCTCCTCTC 
               
               
                   
               
               
                 TTCTCTCTCCCTCTCTCTCTCTTTCTCTTTGCTCATCATCTGTCATTTTGTCCGTTCCTCTCTCTGCGC 
               
               
                   
               
               
                 CCTCAGCGTTCCCCACACTCTCTCGCTTCTCTTTTCCTAGACGTCTTCTTTTTTCATCTTCTTCAGCCT 
               
               
                   
               
               
                 TTTTCGCCATTTTCCATCTCTGTCAATCATTACGGACGACCCCCATTATCGATAAGATCTCCACGGTG 
               
               
                   
               
               
                 GCCGCGAATTCCTGCAGCCCGGGGGATCCCCGGGATTGGCCAAAGGACCCAAAGGTATGTTTCGAA 
               
               
                   
               
               
                 TGATACTAACATAACATAGAACATTTTCAGGAGGACCCTTGGCTAGCGTCGACGGTACCGGGCCCCC 
               
               
                   
               
               
                 CCTCGAGGTCGACGGTATCGATAACCTTCACAGCAGCGCACTCGGTGCCCCGCGCAGGGTCGCGA 
               
               
                   
               
               
                 TG CTGCCCGGTT TGGCACTGTT CCTGCTGGCCGCCTGGACGG CTCGGGCGCT GGATGC 
               
               
                   
               
               
                 AGAATTCCGA CATGACTCAG GATATGAAGT TCATCATCAAAAATTGGTGT TCTTTGCAGA 
               
               
                   
               
               
                 AGATGTGGGT TCAAACAAAG GTGCAATCAT TGGACTCATGGTGGGCGGTG TTGTCATAGC 
               
               
                   
               
               
                 GACAGTGATC GTCATCACCT TGGTGATGCT GAAGAAGAAACAGTACACAT CCATTCATCA 
               
               
                   
               
               
                 TGGTGTGGTG 
               
               
                   
               
               
                 GAGGTTGACG CCGCTGTCAC CCCAGAGGAGCGCCACCTGT CCAAGATGCA GCAGAACGGC 
               
               
                   
               
               
                 TACGAAAATCCAACCTACAATTCTTTGAGCAGATGCAGAACTAGACCCCCGCCACAGCAGCCTCTGA 
               
               
                   
               
               
                 AGTTGGACACGGATCCACTAGTTCTAGAGCGGCCGCCACCGCGGTGGAGCTCCGCATCGGCCGCT 
               
               
                   
               
               
                 GTCATCAGATCGCCATCTCGCGCCCGTGCCTCTGACTTCTAAGTCCAATTACTCTTCAACATCCCTAC 
               
               
                   
               
               
                 ATGCTCTTTCTCCCTGTGCTCCCACCCCCTATTTTTGTTATTATCAAAAAAACTTCTTCTTAATTTCTTT 
               
               
                   
               
               
                 GTTTTTTAGCTTCTTTTAAGTCACCTCTAACAATGAAATTGTGTAGATTCAAAAATAGAATTAATTCGTA 
               
               
                   
               
               
                 ATAAAAAGTCGAAAAAAATTGTGCTCCCTCCCCCCATTAATAATAATTCTATCCCAAAATCTACACAAT 
               
               
                   
               
               
                 GTTCTGTGTACACTTCTTATGTTTTTTTTACTTCTGATAAATTTTTTTTGAAACATCATAGAAAAAACCG 
               
               
                   
               
               
                 CACACAAAATACCTTATCATATGTTACGTTTCAGTTTATGACCGCAATTTTTATTTCTTCGCACGTCTG 
               
               
                   
               
               
                 GGCCTCTCATGACGTCAAATCATGCTCATCGTGAAAAAGTTTTGGAGTATTTTTGGAATTTTTCAATCA 
               
               
                   
               
               
                 AGTGAAAGTTTATGAAATTAATTTTCCTGCTTTTGCTTTTTGGGGGTTTCCCCTATTGTTTGTCAAGAG 
               
               
                   
               
               
                 TTTCGAGGACGGCGTTTTTCTTGCTAAAATCACAAGTATTGATGAGCACGATGCAAGAAAGATCGGA 
               
               
                   
               
               
                 AGAAGGTTTGGGTTTGAGGCTCAGTGGAAGGTGAGTAGAAGTTGATAATTTGAAAGTGGAGTAGTGT 
               
               
                   
               
               
                 CTATGGGGTTTTTGCCTTAAATGACAGAATACATTCCCAATATACCAAACATAACTGTTTCCTACTAGT 
               
               
                   
               
               
                 CGGCCGTACGGGCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACATGCAG 
               
               
                   
               
               
                 CTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGCCCGTCAGGGCGC 
               
               
                   
               
               
                 GTCAGCGGGTGTTGGCGGGTGTCGGGGCTGGCTTAACTATGCGGCATCAGAGCAGATTGTACTGAG 
               
               
                   
               
               
                 AGTGCACCATATGCGGTGTGAAATACCGCACAGATGCGTAAGGAGAAAATACCGCATCAGGCGGCC 
               
               
                   
               
               
                 TTAAGGGCCTCGTGATACGCCTATTTTTATAGGTTAATGTCATGATAATAATGGTTTCTTAGACGTCAG 
               
               
                   
               
               
                 GTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGT 
               
               
                   
               
               
                 ATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGTATGAGTATT 
               
               
                   
               
               
                 CAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAA 
               
               
                   
               
               
                 ACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGAT 
               
               
                   
               
               
                 CTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAA 
               
               
                   
               
               
                 AGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCAT 
               
               
                   
               
               
                 ACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGA 
               
               
                   
               
               
                 CAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGAC 
               
               
                   
               
               
                 AACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTT 
               
               
                   
               
               
                 GATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTA 
               
               
                   
               
               
                 GCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAAT 
               
               
                   
               
               
                 TAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCT 
               
               
                   
               
               
                 GGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGC 
               
               
                   
               
               
                 CAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAAC 
               
               
                   
               
               
                 GAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTA 
               
               
                   
               
               
                 CTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTT 
               
               
                   
               
               
                 GATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAA 
               
               
                   
               
               
                 GATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTG 
               
               
                   
               
               
                 CTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTT 
               
               
                   
               
               
                 TTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGT 
               
               
                   
               
               
                 TAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGT 
               
               
                   
               
               
                 GGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAA 
               
               
                   
               
               
                 GGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACA 
               
               
                   
               
               
                 CCGAACTGAGATACCTACAGCGTGAGCATTGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCG 
               
               
                   
               
               
                 GACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAA 
               
               
                   
               
               
                 ACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGC 
               
               
                   
               
               
                 TCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTT 
               
               
                   
               
               
                 TGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGC 
               
               
                   
               
               
                 CTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGG 
               
               
                   
               
               
                 AAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCT 
               
               
                   
               
               
                 GGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCA 
               
               
                   
               
               
                 CTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGG 
               
               
                   
               
               
                 ATAACAATTTCACACAGGAAACAGCT 
               
             
          
         
       
     
     REFERENCES 
     Baumeister R (1997) Genes &amp; Function 1, 149 
     Daigle I, Li C (1993), 90 (24), 12045 
     Kang, J., Lemaire, H. G., Unterbeck, A., Salbaum J. M., Masters C. L., Grzeschik, K. H., Multhaupt, G., Beyreuther, K., Mueller-Hill, B. (1987) Nature, 325, 733 
     Kenyon, C., Science (1988) 240, 1448 
     Korswagen H. C., Durbin, R. M., Smits, M. T., Plasterk, R. H. A. (1996), 93, 14680 Proc Natl Acad Sci USA 
     Kuwabara, P. E. (1997), Trends in Genetics, 13, 454 
     Levitan D., Doyle T G, Brousseau D., Lee M K. Thinakaran G., Slunt H H., Sisodia S S. Greenwald I. (1996) Proc Natl Acad Sci USA, 93,14940 
     Levitan D, Greenwald I (1995) Nature 377, 351 
     Link C. D. (1995) Proc Natl Acad Sci USA, 92, 9368 
     Mello, C. and Fire, A., Methods in Cell Biology, Academic Press Vol. 48, pp 451, 1995 
     Riddle et al. (1997)  C. elegans  II, Cold Spring Harbor Laboratory Press 
     Rumble, B., Retallack, R., Hilbich, C., Simms, G., Multhaup, G., Martins, R., Hockey, A., Montgomery, P., Beyreuther, K., Masters, C. L., (1989), N. Engl. J. Med., 320, 1446 
     Scheuner, D., Eckman, C., Jensen, M., Song, X., Citron, M., Suzuki, N., Bird, T., Hardy, M., Hutton, W., Kukull, W., Farson, E., Levy-Lahad, E., Vitanen, M., Peskind, E., Poorkaj, P., Schellenberg, G., Tanzi, R., Wasco, W., Lannfeld, D., Selkoe, D., Younkin, S. G. (1996), Nature Medicine, 2, 864 
     Shoji M., Golde T E., Ghiso J., Cheung T T., Estus S., Shaffer L M., Cai X-D., McKay D M., Tintner R., Fraggione B., Younkin S G. (1992) Science 258,126 
     Xiajun Li and Iva Greenwald (1997) Proc Natl Acad Sci USA, 94,12204 
     Yankner, B. A., Caceres, A., Duffy, L. K. (1990) Proc Natl Acad Sci USA, 87, 9020 
     
       
         
           
             11 
           
           
             1 
             303 
             DNA 
             Caenorhabditis elegans 
           
            1
ctggatgcag aattccgaca tgactcagga tatgaagttc atcatcaaaa attggtgttc     60
tttgcagaag atgtgggttc aaacaaaggt gcaatcattg gactcatggt gggcggtgtt    120
gtcatagcga cagtgatcgt catcaccttg gtgatgctga agaagaaaca gtacacatcc    180
attcatcatg gtgtggtgga ggttgacgcc gctgtcaccc cagaggagcg ccacctgtcc    240
aagatgcagc agaacggcta cgaaaatcca acctacaagt tctttgagca gatgcagaac    300
tag                                                                  303
 
           
             2 
             51 
             DNA 
             Caenorhabditis elegans 
           
            2
atgctgcccg gtttggcact gttcctgctg gccgcctgga cggctcgggc g              51
 
           
             3 
             354 
             DNA 
             Caenorhabditis elegans 
           
            3
atgctgcccg gtttggcact gttcctgctg gccgcctgga cggctcgggc gctggatgca     60
gaattccgac atgactcagg atatgaagtt catcatcaaa aattggtgtt ctttgcagaa    120
gatgtgggtt caaacaaagg tgcaatcatt ggactcatgg tgggcggtgt tgtcatagcg    180
acagtgatcg tcatcacctt ggtgatgctg aagaagaaac agtacacatc cattcatcat    240
ggtgtggtgg aggttgacgc cgctgtcacc ccagaggagc gccacctgtc caagatgcag    300
cagaacggct acgaaaatcc aacctacaag ttctttgagc agatgcagaa ctag          354
 
           
             4 
             42 
             DNA 
             Caenorhabditis elegans 
           
            4
gtatgtttcg aatgatacta acataacata gaacattttc ag                        42
 
           
             5 
             495 
             DNA 
             Caenorhabditis elegans 
           
            5
gtatgtttcg aatgatacta acataacata gaacattttc aggaggaccc ttggctagcg     60
tcgacggtac cgggcccccc ctcgaggtcg acggtatcga taaccttcac agcagcgcac    120
tcggtgcccc gcgcagggtc gcgatgctgc ccggtttggc actgttcctg ctggccgcct    180
ggacggctcg ggcgctggat gcagaattcc gaatgactca ggatatgaag tcatcatcaa    240
aaattggtgt tctttgcaga agatgtgggt tcaaacaaag gtgcaatcat tggactcatg    300
gtgggcggtg ttgtcatagc gacagtgatc gtcatcacct tggtgatgct gaagaagaaa    360
cagtacacat ccattcatca tggtgtggtg gaggttgacg ccgctgtcac cccagaggag    420
cgccacctgt ccaagatgca gcagaacggc tacgaaaatc caacctacaa gttctttgag    480
cagatgcaga actag                                                     495
 
           
             6 
             1207 
             DNA 
             Caenorhabditis elegans 
           
            6
aagcttcagt aaaagaagta gaattttata gttttttttc tgtttgaaaa attctcccca     60
tcaatgttct ttcaaataaa tacatcacta atgcaaagta ttctataacc tcatatctaa    120
attcttcaaa atcttaacat atcttatcat tgctttaagt caacgtaaca ttaaaaaaaa    180
tgttttggaa aatgtgtcaa gtctctcaaa attcagtttt ttaaaccact cctatagtcc    240
tatagtccta tagttaccca tgaaatcctt atatattact gtaaaatgtt tcaaaaacca    300
ttggcaaatt gccagaactg aaaatttccg gcaaattggg gaaccggcaa attgccaatt    360
tgctgaattt gccggaaacg gtaattgccg aaagtttttg acacgaaaat ggcaaattgt    420
ggttttaaaa tttttttttt tggaaatttc agaatttcaa ttttaatcgg caaaactgta    480
ggcatcctaa gaatgttcct acatctattt tgaaaagtaa gcgaattaat tctatgaaaa    540
tgtctaaaga aaatggggaa acaatttcaa aaaggcacag tttcaatggt ttccgaatta    600
tactaaatcc ctctaaaaac ttccggcaaa ttgatatccg taaaagagca aatccgcatt    660
tttgccgaaa attaaaattt ccgacaaatc ggcaaaccgg caatttggcg aaatttgccg    720
gaacgattgc cgcccacccc tgttccagag gttcaaactg gtagcaaagc tcaaaatttc    780
tcaaattctc caattttttt ttgaattttg gcagtgtacc aaaatgacat tcagtcatat    840
tggtttatta tagatttatt tagataaaat cctaaatgat tctaccttta aagatgccca    900
ctttaaaagt aatgactcaa acttcaaatt gctctaagat tctattgaat taccatcttt    960
tcctctcatt ttctctcact gtctatttca tcacaaattc atccctctct cctctcttct   1020
ctctccctct ctctctcttt ctctttgctc atcatctgtc attttgtccg ttcctctctc   1080
tgcgccctca gcgttcccca cactctctcg cttctctttt cctagacgtc ttcttttttc   1140
atcttcttca gcctttttcg ccattttcca tctctgtcaa tcattacgga cgacccccat   1200
tatcgat                                                             1207
 
           
             7 
             1773 
             DNA 
             Caenorhabditis elegans 
           
            7
aagcttcagt aaaagaagta gaattttata gttttttttc tgtttgaaaa attctcccca     60
tcaatgttct ttcaaataaa tacatcacta atgcaaagta ttctataacc tcatatctaa    120
attcttcaaa atcttaacat atcttatcat tgctttaagt caacgtaaca ttaaaaaaaa    180
tgttttggaa aatgtgtcaa gtctctcaaa attcagtttt ttaaaccact cctatagtcc    240
tatagtccta tagttaccca tgaaatcctt atatattact gtaaaatgtt tcaaaaacca    300
ttggcaaatt gccagaactg aaaatttccg gcaaattggg gaaccggcaa attgccaatt    360
tgctgaattt gccggaaacg gtaattgccg aaagtttttg acacgaaaat ggcaaattgt    420
ggttttaaaa tttttttttt tggaaatttc agaatttcaa ttttaatcgg caaaactgta    480
ggcatcctaa gaatgttcct acatctattt tgaaaagtaa gcgaattaat tctatgaaaa    540
tgtctaaaga aaatggggaa acaatttcaa aaaggcacag tttcaatggt ttccgaatta    600
tactaaatcc ctctaaaaac ttccggcaaa ttgatatccg taaaagagca aatccgcatt    660
tttgccgaaa attaaaattt ccgacaaatc ggcaaaccgg caatttggcg aaatttgccg    720
gaacgattgc cgcccacccc tgttccagag gttcaaactg gtagcaaagc tcaaaatttc    780
tcaaattctc caattttttt ttgaattttg gcagtgtacc aaaatgacat tcagtcatat    840
tggtttatta tagatttatt tagataaaat cctaaatgat tctaccttta aagatgccca    900
ctttaaaagt aatgactcaa acttcaaatt gctctaagat tctattgaat taccatcttt    960
tcctctcatt ttctctcact gtctatttca tcacaaattc atccctctct cctctcttct   1020
ctctccctct ctctctcttt ctctttgctc atcatctgtc attttgtccg ttcctctctc   1080
tgcgccctca gcgttcccca cactctctcg cttctctttt cctagacgtc ttcttttttc   1140
atcttcttca gcctttttcg ccattttcca tctctgtcaa tcattacgga cgacccccat   1200
tatcgataag atctccacgg tggccgcgaa ttcctgcagc ccgggggatc cccgggattg   1260
gccaaaggac ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga   1320
ggacccttgg ctagcgtcga cggtaccggg ccccccctcg aggtcgacgg tatcgataac   1380
cttcacagca gcgcactcgg tgccccgcgc agggtcgcga tgctgcccgg tttggcactg   1440
ttcctgctgg ccgcctggac ggctcgggcg ctggatgcag aattccgaca tgactcagga   1500
tatgaagttc atcatcaaaa attggtgttc tttgcagaag atgtgggttc aaacaaaggt   1560
gcaatcattg gactcatggt gggcggtgtt gtcatagcga cagtgatcgt catcaccttg   1620
gtgatgctga agaagaaaca gtacacatcc attcatcatg gtgtggtgga ggttgacgcc   1680
gctgtcaccc cagaggagcg ccacctgtcc aagatgcagc agaacggcta cgaaaatcca   1740
acctacaagt tctttgagca gatgcagaac tag                                1773
 
           
             8 
             3344 
             DNA 
             Caenorhabditis elegans 
           
            8
acccccgcca cagcagcctc tgaagttgga cacggatcca ctagttctag agcggccgcc     60
accgcggtgg agctccgcat cggccgctgt catcagatcg ccatctcgcg cccgtgcctc    120
tgacttctaa gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca    180
ccccctattt ttgttattat caaaaaaact tcttcttaat ttctttgttt tttagcttct    240
tttaagtcac ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa    300
aaagtcgaaa aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac    360
acaatgttct gtgtacactt cttatgtttt ttttacttct gataaatttt ttttgaaaca    420
tcatagaaaa aaccgcacac aaaatacctt atcatatgtt acgtttcagt ttatgaccgc    480
aatttttatt tcttcgcacg tctgggcctc tcatgacgtc aaatcatgct catcgtgaaa    540
aagttttgga gtatttttgg aatttttcaa tcaagtgaaa gtttatgaaa ttaattttcc    600
tgcttttgct ttttgggggt ttcccctatt gtttgtcaag agtttcgagg acggcgtttt    660
tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg    720
gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag tagtgtctat    780
ggggtttttg ccttaaatga cagaatacat tcccaatata ccaaacataa ctgtttccta    840
ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct    900
gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac    960
aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg   1020
catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg   1080
taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct atttttatag   1140
gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg   1200
cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga   1260
caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat   1320
ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca   1380
gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc   1440
gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca   1500
atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg   1560
caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca   1620
gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata   1680
accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag   1740
ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg   1800
gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca   1860
acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta   1920
atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct   1980
ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca   2040
gcactgggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg   2100
caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt   2160
ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt   2220
aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac   2280
gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag   2340
atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg   2400
tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca   2460
gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga   2520
actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca   2580
gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc   2640
agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca   2700
ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa   2760
aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc   2820
cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc   2880
gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg   2940
cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat   3000
cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca   3060
gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca   3120
aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg   3180
actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac   3240
cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac   3300
aatttcacac aggaaacagc tatgaccatg attacgccaa gctt                    3344
 
           
             9 
             17 
             PRT 
             Caenorhabditis elegans 
           
            9
Met Leu Pro Gly Leu Ala Leu Phe Leu Leu Ala Ala Trp Thr Ala Arg
  1               5                  10                  15
Ala
 
           
             10 
             117 
             PRT 
             Caenorhabditis elegans 
           
            10
Met Leu Pro Gly Leu Ala Leu Phe Leu Leu Ala Ala Trp Thr Ala Arg
  1               5                  10                  15
Ala Leu Asp Ala Glu Phe Arg His Asp Ser Gly Tyr Glu Val His His
             20                  25                  30
Gln Lys Leu Val Phe Phe Ala Glu Asp Val Gly Ser Asn Lys Gly Ala
         35                  40                  45
Ile Ile Gly Leu Met Val Gly Gly Val Val Ile Ala Thr Val Ile Val
     50                  55                  60
Ile Thr Leu Val Met Leu Lys Lys Lys Gln Tyr Thr Ser Ile His His
 65                  70                  75                  80
Gly Val Val Glu Val Asp Ala Ala Val Thr Pro Glu Glu Arg His Leu
                 85                  90                  95
Ser Lys Met Gln Gln Asn Gly Tyr Glu Asn Pro Thr Tyr Lys Phe Phe
            100                 105                 110
Glu Gln Met Gln Asn
        115
 
           
             11 
             5109 
             DNA 
             Caenorhabditis elegans 
           
            11
atgaccatga ttacgccaag cttcagtaaa agaagtagaa ttttatagtt ttttttctgt     60
ttgaaaaatt ctccccatca atgttctttc aaataaatac atcactaatg caaagtattc    120
tataacctca tatctaaatt cttcaaaatc ttaacatatc ttatcattgc tttaagtcaa    180
cgtaacatta aaaaaaatgt tttggaaaat gtgtcaagtc tctcaaaatt cagtttttta    240
aaccactcct atagtcctat agtcctatag ttacccatga aatccttata tattactgta    300
aaatgtttca aaaaccattg gcaaattgcc agaactgaaa atttccggca aattggggaa    360
ccggcaaatt gccaatttgc tgaatttgcc ggaaacggta attgccgaaa gtttttgaca    420
cgaaaatggc aaattgtggt tttaaaattt ttttttttgg aaatttcaga atttcaattt    480
taatcggcaa aactgtaggc atcctaagaa tgttcctaca tctattttga aaagtaagcg    540
aattaattct atgaaaatgt ctaaagaaaa tggggaaaca atttcaaaaa ggcacagttt    600
caatggtttc cgaattatac taaatccctc taaaaacttc cggcaaattg atatccgtaa    660
aagagcaaat ccgcattttt gccgaaaatt aaaatttccg acaaatcggc aaaccggcaa    720
tttggcgaaa tttgccggaa cgattgccgc ccacccctgt tccagaggtt caaactggta    780
gcaaagctca aaatttctca aattctccaa tttttttttg aattttggca gtgtaccaaa    840
atgacattca gtcatattgg tttattatag atttatttag ataaaatcct aaatgattct    900
acctttaaag atgcccactt taaaagtaat gactcaaact tcaaattgct ctaagattct    960
attgaattac catcttttcc tctcattttc tctcactgtc tatttcatca caaattcatc   1020
cctctctcct ctcttctctc tccctctctc tctctttctc tttgctcatc atctgtcatt   1080
ttgtccgttc ctctctctgc gccctcagcg ttccccacac tctctcgctt ctcttttcct   1140
agacgtcttc ttttttcatc ttcttcagcc tttttcgcca ttttccatct ctgtcaatca   1200
ttacggacga cccccattat cgataagatc tccacggtgg ccgcgaattc ctgcagcccg   1260
ggggatcccc gggattggcc aaaggaccca aaggtatgtt tcgaatgata ctaacataac   1320
atagaacatt ttcaggagga cccttggcta gcgtcgacgg taccgggccc cccctcgagg   1380
tcgacggtat cgataacctt cacagcagcg cactcggtgc cccgcgcagg gtcgcgatgc   1440
tgcccggttt ggcactgttc ctgctggccg cctggacggc tcgggcgctg gatgcagaat   1500
tccgacatga ctcaggatat gaagttcatc atcaaaaatt ggtgttcttt gcagaagatg   1560
tgggttcaaa caaaggtgca atcattggac tcatggtggg cggtgttgtc atagcgacag   1620
tgatcgtcat caccttggtg atgctgaaga agaaacagta cacatccatt catcatggtg   1680
tggtggaggt tgacgccgct gtcaccccag aggagcgcca cctgtccaag atgcagcaga   1740
acggctacga aaatccaacc tacaattctt tgagcagatg cagaactaga cccccgccac   1800
agcagcctct gaagttggac acggatccac tagttctaga gcggccgcca ccgcggtgga   1860
gctccgcatc ggccgctgtc atcagatcgc catctcgcgc ccgtgcctct gacttctaag   1920
tccaattact cttcaacatc cctacatgct ctttctccct gtgctcccac cccctatttt   1980
tgttattatc aaaaaaactt cttcttaatt tctttgtttt tagcttcttt taagtcacct   2040
ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa   2100
aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac aatgttctgt   2160
gtacacttct tatgtttttt ttacttctga taaatttttt ttgaaacatc atagaaaaaa   2220
ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa tttttatttc   2280
ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt   2340
atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg cttttgcttt   2400
ttgggggttt cccctattgt ttgtcaagag tttcgaggac ggcgtttttc ttgctaaaat   2460
cacaagtatt gatgagcacg atgcaagaaa gatcggaaga aggtttgggt ttgaggctca   2520
gtggaaggtg agtagaagtt gataatttga aagtggagta gtgtctatgg ggtttttgcc   2580
ttaaatgaca gaatacattc ccaatatacc aaacataact gtttcctact agtcggccgt   2640
acgggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc   2700
tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg   2760
gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga   2820
ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat   2880
accgcatcag gcggccttaa gggcctcgtg atacgcctat ttttataggt taatgtcatg   2940
ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct   3000
atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga   3060
taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc   3120
cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg   3180
aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc   3240
aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact   3300
tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc   3360
ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag   3420
catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat   3480
aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt   3540
ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa   3600
gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc   3660
aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg   3720
gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt   3780
gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca   3840
gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat   3900
gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca   3960
gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg   4020
atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaact gagttttcgt   4080
tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc   4140
tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc   4200
cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac   4260
caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac   4320
cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt   4380
cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct   4440
gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat   4500
acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt   4560
atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg   4620
cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt   4680
gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt   4740
tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg   4800
tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg   4860
agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc   4920
ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg   4980
gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac   5040
actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag   5100
gaaacagct                                                           5109