Abstract:
The present invention relates to repressors suppressing production of acquired immune deficiency syndrom (AIDS) viral genome (RNA) and methods repressing transcription thereof. The invention, more specifically, relates to fusion proteins repressing AIDS viral genomes (RNA) of a protein selected from group consisting of proteins strongly repressing activities of proteins such as Sp1, NF-kB, proteins repressing transcriptional activities by strongly condensing chromatin, and proteins which are able to bind around AIDS viral promoter; and protein (for example Tat or Tat derivatives) recognizing short RNA strand (TAR) and methods repressing transcription using same. The said fusion proteins dramatically show effect repressing production of HIV-1 genome (RNA) by delivering repressing proteins like Sp1, NF-k B to HIV-1 LTR using protein recognizing short RNA strand as a carrier. The invention shows transcription inhibitory effect of HIV by targeting transcription-repressing fusion proteins to HIV LTR.

Description:
BACKGROUNG OF THE INVENTION  
         [0001]    1. Field of the Invention  
           [0002]    The present invention relates to the repressor for repressing production of acquired immune deficiency syndrome (AIDS) viral genomes (RNA) from proviral or nuclear long terminal repeat (LTR) promoters. More specifically, the present invention relates to fusion proteins for repressing transcription of AIDS viral genomes (RNA), comprising polypeptide sequence selected from the group consisting of: proteins strongly repressing activities of proteins such as Sp1 or NF-κB; proteins repressing transcription by strongly condensing chromatin; protein such as zinc fingers able to bind on AIDS viral promoters; and polypeptide sequence (e.g. Tat protein or Tat derivatives) recognizing short RNA strands (HIV short transcript).  
           [0003]    The fusion proteins have remarkable effects to repress production of the human immunodeficiency virus (HIV)-1 RNA, when proteins strongly repressing activities of proteins such as Sp1 or NF-κB and proteins repressing transcription by strongly condensing chromatin are targeted around the expression control region of HIV-1 LTR promoters by using short RNA strand recognizing proteins (e.g. Tat proteins and mutants thereof) as missile.  
           [0004]    2. Description of the Prior Art  
           [0005]    In general, HIV entry is known to require an binding of the viral envelope protein(GP120) and cell membrane surface receptors. Following binding, the virus fuses with the cell and injects their viral materials. After fusion, viral genome(RNA) is changed to DNA by reverse transcriptase. Additionally, by integrase DNA type viral genome is then integrated into DNA in the host cell where it exists for the life of the cell as a “provinis.” The proviral HIV genome produces the genomic RNA using an expression system of host cell (transcription process), and mass-produces components (proteins, capsids) necessary for proliferation in cytoplasm (translation and protein cleavage, protease function) by using the viral genome. After an assembly process, the HIV genome is then released from the host cell and proliferates. If it is beyond a critical point, 10 billions of new HIVs are proliferated a day on the average. As a result, after infection AIDS is incurable.  
           [0006]    Many anti-AIDS drugs have been developed as antibodies (vaccines) or compounds for inhibiting the above-described processes. Anti-AIDS drugs widely used are inhibitors for reverse transcription process and protease. Although vaccines for inhibiting a process of recognizing host cells has been developed, they have difficulty in suppressing HIV due to rapid mutations of the viral envelope protein. In addition, HIV reverse transcriptase inhibitors and protease inhibitors act as general. HIV suppressors, and various reverse transcriptase inhibitors or protease inhibitors have been developed and marketed in many drug companies. They have effect on inhibition of growth of HIV in the early stage but have some problem such as toxicity and rapid resistance.  
           [0007]    Furthermore, the mechanism study on HIV injection into host cell genomes or therapeutic agent on the process has not been developed. Anti-sense gene therapy and ribozyme gene therapy have been developed since 1997 due to the increasing interest on gene therapy to treat AIDS. A first phase clinical experiment have been in progress since 1997, but the therapy have a deep-seated problem in obtaining viral resistance.  
           [0008]    Many studies on expression control mechanism of growth of viral RNA have demonstrated that transcription control protein in specific host cell is important for RNA expression. Recently, the study have demonstrated that Tat acting with TAK (Tat-associated kinase or PTEF) has great effect on activity of RNA synthetic enzyme. As a result, the recent study places the focus on inhibitors (e.g. Tat analogue protein fragments, TAR analogue RNA segments, TAK inhibitors) for production of RNA by repressing actions of Tat or TAK.  
           [0009]    The above-mentioned therapeutic agents extend life to a certain degree by slowing the process of disease but lose their effect due to rapid resistant viral production. Those therapeutic agents have the basic problem of expression possibility that viral DNA genomes (provirus) existing in the host cell may be expressed into viral RNA because proviral genomes can exist regardless of the above-described agents, vaccines and gene therapy.  
           [0010]    Accordingly, the best way to overcome the problems of the existing agents is to suppress a transcription process that proviral LTR is expressed into genomic RNA. The present invention provides fusion proteins for repressing HIV transcription regulating the expression of AIDS viral RNA. In other words, the fusion proteins of the present invention prevent viral DNA genomes in host cell from being expressed into viral RNA genomes, and then block production of components (RNA genomes, proteins, capsids) necessary for viral proliferation, thereby basically inhibiting proliferation of virus and production of resistant virus. Therefore, the present invention overcomes the problems of the conventional agents and provides an innovating AIDS therapeutic agent.  
         SUMMARY OF THE INVENTION  
         [0011]    In accordance with the present invention, an object of the present invention is to provide a new biological therapeutic agent for basically repressing viral genome(RNA) necessary for proliferation of AIDS and overcoming resistance of virus and a method of repressing HIV transcription to treat AIDS.  
           [0012]    In order to accomplish the above-mentioned object, the present invention provides a fusion protein for repressing HIV transcription, comprising: a polypeptide or compound selected from the group consisting of a) strongly repressing activity of transcription factors such as Sp1 or NF- K B; b) repressing transcriptional activity by condensing chromatin; and c) able to bind the region of promoter (for example, zinc finger); and a polypeptide or compound recognizing RNA strand around expression regulatory regions or the cis-acting regions of viral promoter.  
           [0013]    In the fusion protein in the present invention, the compounds are compounds recognizing specific nucleic acid sequences joining to proteins by enzymatic or chemical methods. The compounds may be small molecules, nucleic acids analogues or oligo-saccharides.  
           [0014]    In the fusion protein of the present invention, the polypeptide or compounds strongly repressing activity of transcription factors such as Sp1 or NF-κB or repressing transcription activity by condensing chromatin, or able to bind transcription regulatory promoter are preferably the polypeptide or compounds selected from the group consisting of: a) POZ-domain proteins; b) HDAC or regions activating transcription inhibition thereof; c) MeCP2 or the analogous MBP-type proteins; d) corepressor proteins selected from the group consisting of polycom family proteins, mSin3A, SMRT and N-CoR; e) DNA binding region polypeptide of Sp1, Sp2, Sp3, Sp4 or NF- K B; and f) proteins(for example; zinc finger) able to bind around HIV promoters.  
           [0015]    Polypeptides or compounds recognizing short transcript around expression regulatory regions or the cis-acting regions of viral promoter are preferably Tat protein shown in SEQ ID NO:1 or 2, its derived polypeptide fragments or mutants thereof.  
           [0016]    It is more preferable that the fusion protein of the present invention is one or more fusion protein selected from the group consisting of proteins shown in SEQ ID NO:3˜10.  
           [0017]    The present invention also provides base sequences of SEQ ID NO:11˜1 8 for coding the polypeptides shown in SEQ ID NO:3˜10 respectively.  
           [0018]    A fusion protein in the present invention, the polypeptides shown in SEQ ID NO:1˜2 are Tat protein mutants consisting of 73 and 72 amino acids, respectively, SEQ ID NO:3 is amino acid sequence of MeCP2-TatdMT(73aa) fusion protein, SEQ ID NO:4 is amino acid sequence of HDAC1-TatdMt(73aa) fusion protein, SEQ ID NO:5 is amino acid sequence of POZ-TatdMt(73aa) fusion protein, SEQ ID NO:6 is amino acid sequence of FBI-1-TatdMt(73aa) fusion protein, SEQ ID NO:7 is amino acid sequence of TatdMt(72aa)-MeCP2 fusion protein, SEQ ID NO:8 is amino acid sequence of TatdMt(72aa)-HDAC1 fusion protein, SEQ ID NO:9 is amino acid sequence of TatdMt(72aa)-POZ fusion protein, and SEQ ID NO:10 is amino acid sequence of TatdMt(72aa)-FBI-1 fusion protein.  
           [0019]    The present invention also provides base sequences shown in SEQ ID NO:11 for coding the MeCP2-TatdMt(73aa) fusion protein, SEQ ID NO:12 is base sequences coding the HDAC1-TatdMt(73aa), SEQ ID NO:13 is base sequences coding the POZ-TatdMt(73aa), SEQ ID NO:14 is base sequences coding the FBI-1-TatdMt(73aa), SEQ ID NO:15 is base sequences coding the TatdMt(72aa)-MeCP2, SEQ ID NO:16 is base sequences coding the TatdMt(72aa)-HDAc1, SEQ ID NO:17 is base sequences coding TatdMt(72aa)-POZ, and SEQ ID NO:18 is base sequences coding TatdMt(72aa)-FBI-1.  
           [0020]    The present invention provides compositions for suppressing proliferation of HIV including a portion or the whole of the fusion proteins.  
           [0021]    The present invention provides a portion or the whole of base sequences of one or more recombinant vector selected from the group consisting of pcDNA3.0-TatWt, pcDNA3.0-TatMt, pcDNA3.0-FBI-1, pcDNA3.0-MeCP2-TatWt, pcDNA3.0HDAC1-TatWt, pcDNA3.0FBI-1-TatWt, pcDNA3.0-POZ-TatWt, pcDNA3.0TarWt-MeCP2, pcDNA3.0TatWt-HDAC1, pcDNA3.0TatWt-FBI-1, pcDNA3.0TatWt-POZ, pcDNA3.0-MeCP2-TatdMt, pcDNA3.0HDAC1-TatdMt, pcDNA3.0FBI-1-TatdMt, pcDNA3.0-POZ-TatdMt, pcDNA3.0TatdMt-Mecp2, pcDNA3.0TatdMt-HDAC1, pcDNA3.0TatdMt-FBI-1 and pcDNA3.0TatdMt-POZ comprising genes coding the above-mentioned fusion proteins.  
           [0022]    The present invention provides a method for suppressing transcription of viral genome(RNA) by targeting proteins or materials repressing transcription by using protein or material having binding activity to HIV short transcripts or promoter regulatory regions (cis-acting elements). 
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0023]    [0023]FIG. 1 is a picture illustrating the transcription control of viral genome(RNA) in HIV LTR region. RNA synthesis at the low level is mainly determined by Sp1, NF-κB and TATA box. Defective Sp1-associated GC-Box or TATA box prevents transcription. If transcription is happened once, HIV short strands (short transcripts) exist around LTR promoters. If Tat proteins bind TAR regions, PTEF (positive transcription elongation factor) binds and then CDK9 (cyclin dependent kinase 9) strongly phosphorylates CTD (C Terminal domain) of polymerase II (Pol II). As a result, long viral RNA is formed, thereby rapidly replicating HIV.  
         [0024]    [0024]FIG. 2 shows the construct of repressor-TatWt (consisting of wild type 86 amino acid of Tat protein) fusion proteins(a), and (b) shows the analysis result of transient expression in CV-1 cells. The fusion proteins fused with TatWt may not inhibit viral genome expression in HIV-LTR at the level of transcription, but the fusion proteins may be targeted into HIV LTR promoters by TatWt part, thereby enhancing expression by stimulating transcription elongation.  
         [0025]    [0025]FIG. 3 shows the construct(a) of repressor-TatdMt (mutants consisting of 72 or 73 amino acids wherein 2 amino acid sequences of Tat proteins are mutated), (b) shows the result of transient expression in CV-1 cells, and (c) shows the result of expression in HeLa cells wherein HIV-1 LTR promoters are inserted into genomes like provirus state. The fusion proteins fused with TatdMt strongly inhibit transcription of viral genome in HIV-LTR in the presence of 300 ng of TatWt expression plasmid, thereby reduce the expression by the low level in the absence of Tat. The result reveals that transcription in HIV-LTR like provirus state in HeLa cell strongly inhibit by targeting fusion protein to TAR region by Tat.  
         [0026]    [0026]FIG. 4 shows function of TatWt and FBI-1, FBI-1-TatdMt, TatdMt in HIV-1 LTR promoters. FBI-1 itself does not show transcription inhibitory function. TatdMt shows only competitive inhibitory function. When the same amount of FBI-1 and TatdMt is provided, about 50% of inhibitory function is shown(b8). But FBI-1-TatdMt shows inhibitory function at 1 ng, and 50% inhibitory function at 3 ng. FBI-1-TatdMt shows the same inhibitory function at 81 ng as that of the absence of TatWt, thereby completely inhibiting transcription by TatWt. However, FBI-1-TatdMt shows a lower level of expression suppression at 243 ng than in the absence of TatWt. 
     
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS  
       [0027]    The present invention will be explained in terms of exemplary embodiments described in detail with reference to the accompanying drawings, which are given only by way of illustration and thus are not limitative of the present invention.  
         [0028]    Regulation of synthesizing HIV RNA is complicated, and it requires interaction with cis-acting elements in various viral LTRs, viral transactivators and cellular proteins. This interaction controls the basic level of RNA transcription, and induces expression of high amount of viral genes. Transcription in HIV-LTR promoters is mainly regulated by cellular protein Sp1 transcription factors for recognizing adjacent promoters in the absence of viral protein Tat.  
         [0029]    Various signals inducing activation of NF-κB activates transcription by interacting with cis-acting elements existing in upstream of Sp1-associated GC-Box. Other proteins (cofactors) promote transcription and replication in cells having latent HIV provirus by regulating activity of proteins such as Sp1 or NF-κB.  
         [0030]    Tat proteins which HIV codes activate transcription by increasing the initiation or elongation of transcription, and they are important for replication. Tat requires TAR, which is a cis-acting RNA element existing in the 5′ end of viral transcripts. Tat also highly promotes production of viral RNA having TAR stem-loop RNA structure in the 5′ end of all HIV transcripts.  
         [0031]    As a result of the recent study, Sp1 is repressed by interaction with HDAC and POZ-domain through zinc-finger DNA binding domain, and also by an interaction with MeCP2. The study has demonstrated that a POZ-domain transcription factor referred to as FBI-1 inhibits HIV transcription by interacting with Sp1 and Tat and binding to an IST region (short strand inductive region) of HIV-LTR. These proteins interact with corepressor proteins such as mSin3A, SMRT/N-CoR and HAD for inhibiting transcription by condensing chromatin. Accordingly, if these proteins are targeted in HIV-1 LTR, the activity of transcription by Sp1 and Tat may be regulated by blocking activity of Sp1, condensing chromatin around HIV promoters and targeting polypeptides able to bind HIV promoter region.  
         [0032]    Expression plasmid fused transcription inhibitory protein with TatWt is prepared in order to target transcription inhibitory proteins to core promoter regions. Inventors in the present invention make TatWt fusion proteins not inhibit transcription in HIV LTR (promoters) in cell and TatWt fusion proteins function as transcription activation factor of viral RNA by TatWt part of fusion proteins. Function of transcription stimulatory factor of transcription inhibitory proteins fused with TatWt is regarded that TatWt potently functions in transcription elongation step enough to compensate with function of inhibitors inhibiting transcription initiation. However, although fusion proteins consist of two different kinds of proteins, they may be targeted to TAR by TatWt part. As a result, Tat proteins may be used as a targeting means to a core LTR promoter region.  
         [0033]    Accordingly, the present invention uses Tat protein mutant (TatdMt:TatK28A&amp;K50A) strongly binding to TAR but lacking an interaction with TAK (or referred to as ‘PTEF’) which is an important cellular factor in transcription activation by Tat.  
         [0034]    The longest Tat of HIV consists of 101 amino acids. Because there are many diverse mutants, it is difficult to distinguish which kind of Tats a wild type. The present invention uses Tat proteins wherein Lys-28 and Lys-50 are substituted with alanine, and uses a Tat polypeptide consisting of 72 or 73 amino acids. However, fragments of the Tat polypeptide or other Tat mutants besides the above-mentioned Tat have the similar function as described above.  
         [0035]    TatdMt fusion proteins dramatically inhibit transcription of HIV genomes in simian CV-1 cells even when excessive Tat (300 ng of expression plasmid) is expressed.  
         [0036]    This experimental result is an epoch-making discovery of a protein having a dominant-negative form. Most Tat fusion proteins function regardless of their directions binding to TatdMt. Particularly HDAC-TatdMt, FBI-1-TatdMt fusion proteins strongly inhibit the transcription (see FIG. 3 b ). The fusion proteins strongly act as inhibitors in HeLa cells wherein HIV-1 LTR-chloramphenicol acetyl transferase gene is inserted into human genomes. Although Tat protein itself activates transcription by 46 times, MeCP2, HDAC, POZ-, or FBI-1-TatdMt fusion proteins strongly inhibit the transcription activation by TatWt even under 300 ng of TatWt over expression plasmids condition. HDAC-TatdMt and FBI-1-TatdMt among the fusion proteins completely inhibit the transcription activation by Tat in HeLa cells too. This result shows that the fusion proteins may repress the transcription of HIV inserted into human genomes as provirus state, in consequence effectively inhibit the proliferation of HIV.  
         [0037]    Because the fusion proteins of the present invention repress transcription in HIV LTR promoter and RNA necessary to produce all components for viral replication is not produced, it necessarily follows that the fusion proteins repress expression of viral proteins and replication of HIV.  
       Example 1  
     Preparation of TatWt Plasmid  
       [0038]    HIV-1 LTR CAT fusion plasmid (pUC3R-III CAT) was prepared by cloning about 720 bp of HIV-LTR to pCAT-Basic plasmid (Promega Co.). HIV-1 LTR Luciferase fusion plasmid (pUC3R-III-Luc) was prepared by cloning 720 bp segments of pUC3R-III CAT digested with Xho I HidIII restriction enzymes to pGL3-Basic plasmid (Xho I HidIII, Promega Co.).  
         [0039]    The fusion proteins of the present invention fused with Tat (consisting of 86 amino acids), TatdMt (consisting of 73 amino acids), HDAC1, MeCP2, FBI-1, POZ-domain and TatWt or mutant TatdMt (TatK28AK50A) were prepared by amplifying the corresponding gene by a PCR method, and then cloning the genes to a mammalian expression vector pcDNA3.0(Inivtorgen). In order to prepare pcDNA3.0 TatWt(86 amino acids), The genes were amplified by a PCR method using pET-15b-TATWt (86aa, provided by phD. Park Jinseo in Hallym Univ., Korea, HIVHBX2R type) as a template and  
                                       5′ primer:               GATCGAATTCATGGAGCCAGTACCTAGACTAGAGCCC,                       3′ primer:           GATCTCTAGATCATTCCTTCGGGCCTGTCGGGTCCCCTC.          
 
         [0040]    The reaction conditions were as follows: template denaturation at 94° C. for 3 minutes, 30 cycles of amplification reaction (94° C. 30 sec.; 60° C. 1 min.; 72° C. 30 sec.), and then post-amplification reaction at 72° C. for 3 minutes. After PCR products and expression vector pcDNA3.0 were digested with two restriction enzyme EcoR1 and Xba1, the digested products were ligated using T4 DNA ligase, and then introduced into  E. coli  DH5a by a transformation method and pcDNA3.0 TatWt(86 amino acids) plasmid was prepared by an alkali lysis method.  
         [0041]    Next, in order to prepare pcDNA3.0 TatdMt(73aa), methods similar to the above mentioned methods were used. However, pcDNA3.0TatdMt was prepared by a PCR amplification using HIV Tat gene (Kiernan et al., EMBO J. 18:6106-6118, 1999) as a template and 5′ primer: GAT CGA ATT CAT GGA GCC AGT AAA TCC TAG CCT AG, 3′ Primer: GATCTCTAGATCAGCTTTGATAGAGAAACTTGATG (containing stop codon). In order to prepare HDAC, MeCP2, POZ-, or FBI-1 fusion proteins of X-TatdMT, non-Tat portions were prepared by amplifying the corresponding human cDNA using PCR method and then by cloning the amplified genes to pcDNA3.0 TatdMt. The Reaction conditions were as follows: template denaturation at 95° C. for 3 minutes, 30 cycles of amplification (95° C. 30 sec.; 62° C. 1 min.; 72° C. 7 min.) and post-amplification reaction at 72° C. for 3 minutes, and PCR reaction was carried out using the following PCR primers:  
                                       MeCP2               5′ primer:           GATCGGATCCACCATGGTAGCTGGGATGTTAGGGCTCAG                       3′ primer:           GATCGAATTCGCTAACTCTCTCGGTCACGGGCGTCCG                       HCAC1           5′ primer:           GATCAAGCTTACCATGGCGCAGACGCAGGGCACCCGGAGG                       3′ primer:           GATCGAATTCGGCCAACTTGACCTCCTCCTTGACCCCTTTG                       POZ-domain           5′ primer:           GATCAAGCTTACCATGGCCGGCGGCGTGGACGGCCCCATC                       3′ primer:           GATCGAATTCCTGCCGGTCCAGGAGGTCGGCGCACACG                       FBI-1           5′ primer:           GATCAAGCTTACCATGGCCGGCGGCGTGGACGGCCCCATC                       3′ primer:           GATCAGATTCGGCGAGTCCGGCTGTGAAGTT.          
 
         [0042]    The amplified products were digested with BamH1-EcoR1 (MeCP2) or HindIII-EcoR1(HDAC1, POZ, FBI-1), and then cloned to pcDNA3.0 TatdMt/BamH1-EcoR1 or pcDNA3.0 TatdMt/HindIII-EcoR1.  
         [0043]    In order to prepare HDAC, MeCP2, POZ-, FBI-1 fusion proteins of pcDNA3.0 TatdMt-X, methods similar to the above mentioned methods were used. However, pcDNA3.0 TatdMt(73aa) was prepared by PCR amplification using HIV Tat gene (Kiernan et al., EMBO J. 18: 6106-6118, 1999) as a template and  
                           5′ primer: GATCGGATCCACCATGGACGGAGTAAATCCTAGCCTAG                   3′ primer: GATCGAATTCGGGCTTTGATAGAGAAACTTGATG.          
 
         [0044]    Non-Tat portions of pcDNA3.0 TatdMt-x family were prepared by amplifying the corresponding human cDNA in the same condition the above mentioned and by digesting the amplified products with EcoR1-Xba1 and by cloning the digested products to pcDNA3.0 TatdMt/EcoR1-Xba1. Primer sets used in amplification reaction were as follows:  
                                       MeCP2               5′ primer: GATCGAATTCATGGTAGCTGGGATGTTAGGGCTCA                       3′ primer: GATCTCTAGATCAGCTAACTCTCTCGGTCACGGGC                       HDAC1           5′ primer: GATCGAATTCATGGCGCAGACGCAGGGCACCCGGA                       3′ primer: GATCTCTAGATCAGGCCAACTTGACCTCCTCCTTG                       POZ-domain           5′ primer: GATCGAATTCATGGCCGGCGGCGGCGTGGACGGCC                       3′ primer: GATCTCTAGATCACTGCCGGTCCAGGAGGTCGGCG                       FBI-1           5′ primer: GATCGAATTCATGGCCGGCGGCGGCGTGGACGGCC                       3′ primer: GATCTCTAGATCAGGCGAGTCCGGCTGTGAAGTT.          
 
       Example 2  
     Transient Expression Assay  
       [0045]    CV-1 cells were cultured in a DMEM culture medium with 10% FBS. When the cells grew enough to occupy 50-60% of bottom area in culture vessel, the mixtures consisting of 0.6 μg of pHIV-LTR-Luciferase plasmid, pCMV-β galactosidase plasmid, TatWt, and a mammalian expression pcDNA3.0 plasmid selected from the group consisting of HDAC1-TatdMt, MeCP2-TatdMt, FBI-1-TatdMt, POZ-domian-TatdMt, TatdMt-HDAC1, TatdMt-MeCP2, TatdMt-FBI-1, and TatdMt-POZ-domain were introduced into cell using a lipopectamin plus reagent(Gibco-BRL).  
         [0046]    In order to perform an experiment for repressing genome expressions in HeLa cells which HIV-LTR-CAT was inserted into genome, the mixtures consisting of 300 ng of Tat expression plasmid and 300 ng of various fusion proteins expression plasmid of the present invention were introduced into cells were cultured in DMEM culture medium with 10% FBS using the lipopectamin plus reagent (Gibco-BRL). The cells were cultured for 24 hours, and the expression of reporter gene was analyzed. The efficiency for introducing plasmid into cell and the deviation for recovering cell extracts were standardized using activity of simultaneously introduced and expressed β-galactosidase. The results of the present invention are shown in FIGS.  2 ˜ 4 .  
         [0047]    In FIG. 2, the fusion proteins fused with TatWt may not inhibit genome expression in HIV-LTR at transcriptional level, but the fusion proteins may be targeted into HIV LTR promoter region by TatWt part, thereby enhancing expression by promoting transcription elongation.  
         [0048]    In FIG. 3, the fusion proteins fused with TatdMt (mutants consisting of 72 or 73 amino acids wherein 2 amino acid sequences of Tat proteins are mutated) potently inhibit genome transcription in HIV-LTR in the presence of 300 ng of TatWt expression plasmid, thereby reduce the expression by the low level in the absence of Tat. The result reveals that transcription in HIV-LTR like provirus state in HeLa cell strongly inhibit by targeting fusion protein to TAR region by Tat.  
         [0049]    In FIG. 4, FBI-1 itself does not show transcription inhibitory function. TatdMt shows only competitive inhibition that competitively binds to the same site (TAR). When the same amount of FBI-1 and TatdMt is provided, about 50% of inhibitory function is shown(b8). But FBI-1-TatdMt shows inhibitory function at 1 ng, and 50% inhibitory function at 3 ng. FBI-1-TatdMt shows the same inhibitory function at 81 ng as that of the absence of TatWt, thereby completely inhibiting transcription by TatWt. However, FBI-1-TatdMt shows a lower level of expression suppression at 243 ng than in the absence of TatWt.  
         [0050]    Within cell nucleus, host cellular proteins such as Sp1, NF-κB act upon HIV transcription regulatory regions. As a result, viral RNA of short RNA strand (short transcript) is produced and it stays around the regions (representing RNA strands binding to polymerase II). If viral protein such as Tat binds to TAR region, RNA synthesis is highly promoted. As a result, a large amount of viral genome (RNA) is produced, and protein components necessary for viral proliferation are produced by using the resulting viral genome(RNA). Accordingly, the most effective method to inhibit viral proliferation is to regulate function of Sp1, NF- k B, Tat.  
         [0051]    The present invention provides the method to potently inhibit production of viral RNA (transcription process) in viral LTR, when protein for recognizing HIV short transcript regions (e.g. TAR) is targeted to HIV transcription regulatory promoter region, by fusing the protein with the protein selected from the group consisting of proteins repressing transcription factor like Sp1, NF-κB; corepressor or proteins interacting with corepressor; HDAC, or proteins interacting with HDAC repressing transcriptional activity by strongly condensing chromatin; and proteins such as zinc finger having binding activity to viral promoter region.  
         [0052]    The present invention is the first study showing the method of potently inhibiting transcription activity in HIV LTR by targeting transcription inhibitory protein groups to HIV-LTR. The present invention also basically inhibits production of components (genomes, proteins) necessary for viral proliferation by repressing expression of viral RNA genome from DNA type viral LTR (promoter) existing as proviral state in host cellular genome. Accordingly, the present invention may basically inhibit the proliferation of virus and production of resistant virus. Particularly, HDAC-TatdMt and FBI-1-TatdMt fusion proteins completely block the process of producing viral genome (RNA).  
         [0053]    Therefore, the present invention to overcome the problems of the conventional drugs as AIDS therapeutic agents is the effective protein or gene therapeutic agent to treat AIDS.  
     
       
       
         1 
         
           
             40  
           
           
             1  
             73  
             PRT  
             Artificial Sequence  
             
               73 amino acid Tat mutant  
             
           
            1 

Met Glu Pro Val Asn Pro Ser Leu Glu Pro Trp Lys His Pro Gly Ser 
  1               5                  10                  15 

Gln Pro Lys Thr Ala Cys Thr Asn Cys Tyr Cys Ala Lys Cys Cys Phe 
             20                  25                  30 

His Cys Gln Val Cys Phe Ile Thr Lys Ala Leu Gly Ile Ser Tyr Gly 
         35                  40                  45 

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

His Gln Val Ser Leu Ser Lys Leu Ile 
 65                  70 

 
           
             2  
             72  
             PRT  
             Artificial Sequence  
             
               72 amino acid Tat mutant  
             
           
            2 

Met Glu Pro Val Asn Pro Ser Leu Glu Pro Trp Lys His Pro Gly Ser 
  1               5                  10                  15 

Gln Pro Lys Thr Ala Cys Thr Asn Cys Tyr Cys Ala Lys Cys Cys Phe 
             20                  25                  30 

His Cys Gln Val Cys Phe Ile Thr Lys Ala Leu Gly Ile Ser Tyr Gly 
         35                  40                  45 

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

His Gln Val Ser Leu Ser Lys Leu 
 65                  70 

 
           
             3  
             561  
             PRT  
             Artificial Sequence  
             
               MeCP2-Tat dMt fusion protein  
             
           
            3 

Met Val Ala Gly Met Leu Gly Leu Arg Glu Glu Lys Ser Glu Asp Gln 
  1               5                  10                  15 

Asp Leu Gln Gly Leu Lys Asp Lys Pro Leu Lys Phe Lys Lys Val Lys 
             20                  25                  30 

Lys Asp Lys Lys Glu Glu Lys Glu Gly Lys His Glu Pro Val Gln Pro 
         35                  40                  45 

Ser Ala His His Ser Ala Glu Pro Ala Glu Ala Gly Lys Ala Glu Thr 
     50                  55                  60 

Ser Glu Gly Ser Gly Ser Ala Pro Ala Val Pro Glu Ala Ser Ala Ser 
 65                  70                  75                  80 

Pro Lys Gln Arg Arg Ser Ile Ile Arg Asp Arg Gly Pro Met Tyr Asp 
                 85                  90                  95 

Asp Pro Thr Leu Pro Glu Gly Trp Thr Arg Lys Leu Lys Gln Arg Lys 
            100                 105                 110 

Ser Gly Arg Ser Ala Gly Lys Tyr Asp Val Tyr Leu Ile Asn Pro Gln 
        115                 120                 125 

Gly Lys Ala Phe Arg Ser Lys Val Glu Leu Ile Ala Tyr Phe Glu Lys 
    130                 135                 140 

Val Gly Asp Thr Ser Leu Asp Pro Asn Asp Phe Asp Phe Thr Val Thr 
145                 150                 155                 160 

Gly Arg Gly Ser Pro Ser Arg Arg Glu Gln Lys Pro Pro Lys Lys Pro 
                165                 170                 175 

Lys Ser Pro Lys Ala Pro Gly Thr Gly Arg Gly Arg Gly Arg Pro Lys 
            180                 185                 190 

Gly Ser Gly Thr Thr Arg Pro Lys Ala Ala Thr Ser Glu Gly Val Gln 
        195                 200                 205 

Val Lys Arg Val Leu Glu Lys Ser Pro Gly Lys Leu Leu Val Lys Met 
    210                 215                 220 

Pro Phe Gln Thr Ser Pro Gly Gly Lys Ala Glu Gly Gly Gly Ala Thr 
225                 230                 235                 240 

Thr Ser Thr Gln Val Met Val Ile Lys Arg Pro Gly Arg Lys Arg Lys 
                245                 250                 255 

Ala Glu Ala Asp Pro Gln Ala Ile Pro Lys Lys Arg Gly Arg Lys Pro 
            260                 265                 270 

Gly Ser Val Val Ala Ala Ala Ala Ala Glu Ala Lys Lys Lys Ala Val 
        275                 280                 285 

Lys Glu Ser Ser Ile Arg Ser Val Gln Glu Thr Val Leu Pro Ile Lys 
    290                 295                 300 

Lys Arg Lys Thr Arg Glu Thr Val Ser Ile Glu Val Lys Glu Val Val 
305                 310                 315                 320 

Lys Pro Leu Leu Val Ser Thr Leu Gly Glu Lys Ser Gly Lys Gly Leu 
                325                 330                 335 

Lys Thr Cys Lys Ser Pro Gly Arg Lys Ser Lys Glu Ser Ser Pro Lys 
            340                 345                 350 

Gly Arg Ser Ser Ser Ala Ser Ser Pro Pro Lys Lys Glu His His His 
        355                 360                 365 

His His His His Ser Glu Ser Pro Lys Ala Pro Val Pro Leu Leu Pro 
    370                 375                 380 

Pro Leu Pro Pro Pro Pro Pro Glu Pro Glu Ser Ser Glu Asp Pro Thr 
385                 390                 395                 400 

Ser Pro Pro Glu Pro Gln Asp Leu Ser Ser Ser Val Cys Lys Glu Glu 
                405                 410                 415 

Lys Met Pro Arg Gly Gly Ser Leu Glu Ser Asp Gly Cys Pro Lys Glu 
            420                 425                 430 

Pro Ala Lys Thr Gln Pro Ala Val Ala Thr Ala Ala Thr Ala Ala Glu 
        435                 440                 445 

Lys Tyr Lys His Arg Gly Glu Gly Glu Arg Lys Asp Ile Val Ser Ser 
    450                 455                 460 

Ser Met Pro Arg Pro Asn Arg Glu Glu Pro Val Asp Ser Arg Thr Pro 
465                 470                 475                 480 

Val Thr Glu Arg Val Ser Glu Phe Met Glu Pro Val Asn Pro Ser Leu 
                485                 490                 495 

Glu Pro Trp Lys His Pro Gly Ser Gln Pro Lys Thr Ala Cys Thr Asn 
            500                 505                 510 

Cys Tyr Cys Ala Lys Cys Cys Phe His Cys Gln Val Cys Phe Ile Thr 
        515                 520                 525 

Lys Ala Leu Gly Ile Ser Tyr Gly Arg Ala Lys Arg Arg Gln Arg Arg 
    530                 535                 540 

Arg Pro Pro Gln Gly Ser Gln Thr His Gln Val Ser Leu Ser Lys Leu 
545                 550                 555                 560 

Ile 

 
           
             4  
             557  
             PRT  
             Artificial Sequence  
             
               HDAC1-TAT dMt fusion protein  
             
           
            4 

Met Ala Gln Thr Gln Gly Thr Arg Arg Lys Val Cys Tyr Tyr Tyr Asp 
  1               5                  10                  15 

Gly Asp Val Gly Asn Tyr Tyr Tyr Gly Gln Gly His Pro Met Lys Pro 
             20                  25                  30 

His Arg Ile Arg Met Thr His Asn Leu Leu Leu Asn Tyr Gly Leu Tyr 
         35                  40                  45 

Arg Lys Met Glu Ile Tyr Arg Pro His Lys Ala Asn Ala Glu Glu Met 
     50                  55                  60 

Thr Lys Tyr His Ser Asp Asp Tyr Ile Lys Phe Leu Arg Ser Ile Arg 
 65                  70                  75                  80 

Pro Asp Asn Met Ser Glu Tyr Ser Lys Gln Met Gln Arg Phe Asn Val 
                 85                  90                  95 

Gly Glu Asp Cys Pro Val Phe Asp Gly Leu Phe Glu Phe Cys Gln Leu 
            100                 105                 110 

Ser Thr Gly Gly Ser Val Ala Ser Ala Val Lys Leu Asn Lys Gln Gln 
        115                 120                 125 

Thr Asp Ile Ala Val Asn Trp Ala Gly Gly Leu His His Ala Lys Lys 
    130                 135                 140 

Ser Glu Ala Ser Gly Phe Cys Tyr Val Asn Asp Ile Val Leu Ala Ile 
145                 150                 155                 160 

Leu Glu Leu Leu Lys Tyr His Gln Arg Val Leu Tyr Ile Asp Ile Asp 
                165                 170                 175 

Ile His His Gly Asp Gly Val Glu Glu Ala Phe Tyr Thr Thr Asp Arg 
            180                 185                 190 

Val Met Thr Val Ser Phe His Lys Tyr Gly Glu Tyr Phe Pro Gly Thr 
        195                 200                 205 

Gly Asp Leu Arg Asp Ile Gly Ala Gly Lys Gly Lys Tyr Tyr Ala Val 
    210                 215                 220 

Asn Tyr Pro Leu Arg Asp Gly Ile Asp Asp Glu Ser Tyr Glu Ala Ile 
225                 230                 235                 240 

Phe Lys Pro Val Met Ser Lys Val Met Glu Met Phe Gln Pro Ser Ala 
                245                 250                 255 

Val Val Leu Gln Cys Gly Ser Asp Ser Leu Ser Gly Asp Arg Leu Gly 
            260                 265                 270 

Cys Phe Asn Leu Thr Ile Lys Gly His Ala Lys Cys Val Glu Phe Val 
        275                 280                 285 

Lys Ser Phe Asn Leu Pro Met Leu Met Leu Gly Gly Gly Gly Tyr Thr 
    290                 295                 300 

Ile Arg Asn Val Ala Arg Cys Trp Thr Tyr Glu Thr Ala Val Ala Leu 
305                 310                 315                 320 

Asp Thr Glu Ile Pro Asn Glu Leu Pro Tyr Asn Asp Tyr Phe Glu Tyr 
                325                 330                 335 

Phe Gly Pro Asp Phe Lys Leu His Ile Ser Pro Ser Asn Met Thr Asn 
            340                 345                 350 

Gln Asn Thr Asn Glu Tyr Leu Glu Lys Ile Lys Gln Arg Leu Phe Glu 
        355                 360                 365 

Asn Leu Arg Met Leu Pro His Ala Pro Gly Val Gln Met Gln Ala Ile 
    370                 375                 380 

Pro Glu Asp Ala Ile Pro Glu Glu Ser Gly Asp Glu Asp Glu Asp Asp 
385                 390                 395                 400 

Pro Asp Lys Arg Ile Ser Ile Cys Ser Ser Asp Lys Arg Ile Ala Cys 
                405                 410                 415 

Glu Glu Glu Phe Ser Asp Ser Glu Glu Glu Gly Glu Gly Gly Arg Lys 
            420                 425                 430 

Asn Ser Ser Asn Phe Lys Lys Ala Lys Arg Val Lys Thr Glu Asp Glu 
        435                 440                 445 

Lys Glu Lys Asp Pro Glu Glu Lys Lys Glu Val Thr Glu Glu Glu Lys 
    450                 455                 460 

Thr Lys Glu Glu Lys Pro Glu Ala Lys Gly Val Lys Glu Glu Val Lys 
465                 470                 475                 480 

Leu Ala Glu Phe Met Glu Pro Val Asn Pro Ser Leu Glu Pro Trp Lys 
                485                 490                 495 

His Pro Gly Ser Gln Pro Lys Thr Ala Cys Thr Asn Cys Tyr Cys Ala 
            500                 505                 510 

Lys Cys Cys Phe His Cys Gln Val Cys Phe Ile Thr Lys Ala Leu Gly 
        515                 520                 525 

Ile Ser Tyr Gly Arg Ala Lys Arg Arg Gln Arg Arg Arg Pro Pro Gln 
    530                 535                 540 

Gly Ser Gln Thr His Gln Val Ser Leu Ser Lys Leu Ile 
545                 550                 555 

 
           
             5  
             205  
             PRT  
             Artificial Sequence  
             
               POZ-TAT dMt fusion protein  
             
           
            5 

Met Ala Gly Gly Val Asp Gly Pro Ile Gly Ile Pro Phe Pro Asp His 
  1               5                  10                  15 

Ser Ser Asp Ile Leu Ser Gly Leu Asn Glu Gln Arg Thr Gln Gly Leu 
             20                  25                  30 

Leu Cys Asp Val Val Ile Leu Val Glu Gly Arg Glu Phe Pro Thr His 
         35                  40                  45 

Arg Ser Val Leu Ala Ala Cys Ser Gln Tyr Phe Lys Lys Leu Phe Thr 
     50                  55                  60 

Ser Gly Ala Val Val Asp Gln Gln Asn Val Tyr Glu Ile Asp Phe Val 
 65                  70                  75                  80 

Ser Ala Glu Ala Leu Thr Ala Leu Met Asp Phe Ala Tyr Thr Ala Thr 
                 85                  90                  95 

Leu Thr Val Ser Thr Ala Asn Val Gly Asp Ile Leu Ser Ala Ala Arg 
            100                 105                 110 

Leu Leu Glu Ile Pro Ala Val Ser His Val Cys Ala Asp Leu Leu Asp 
        115                 120                 125 

Arg Gln Glu Phe Met Glu Pro Val Asn Pro Ser Leu Glu Pro Trp Lys 
    130                 135                 140 

His Pro Gly Ser Gln Pro Lys Thr Ala Cys Thr Asn Cys Tyr Cys Ala 
145                 150                 155                 160 

Lys Cys Cys Phe His Cys Gln Val Cys Phe Ile Thr Lys Ala Leu Gly 
                165                 170                 175 

Ile Ser Tyr Gly Arg Ala Lys Arg Arg Gln Arg Arg Arg Pro Pro Gln 
            180                 185                 190 

Gly Ser Gln Thr His Gln Val Ser Leu Ser Lys Leu Ile 
        195                 200                 205 

 
           
             6  
             659  
             PRT  
             Artificial Sequence  
             
               FBI-1-TAT dMt fusion protein  
             
           
            6 

Met Ala Gly Gly Val Asp Gly Pro Ile Gly Ile Pro Phe Pro Asp His 
  1               5                  10                  15 

Ser Ser Asp Ile Leu Ser Gly Leu Asn Glu Gln Arg Thr Gln Gly Leu 
             20                  25                  30 

Leu Cys Asp Val Val Ile Leu Val Glu Gly Arg Glu Phe Pro Thr His 
         35                  40                  45 

Arg Ser Val Leu Ala Ala Cys Ser Gln Tyr Phe Lys Lys Leu Phe Thr 
     50                  55                  60 

Ser Gly Ala Val Val Asp Gln Gln Asn Val Tyr Glu Ile Asp Phe Val 
 65                  70                  75                  80 

Ser Ala Glu Ala Leu Thr Ala Leu Met Asp Phe Ala Tyr Thr Ala Thr 
                 85                  90                  95 

Leu Thr Val Ser Thr Ala Asn Val Gly Asp Ile Leu Ser Ala Ala Arg 
            100                 105                 110 

Leu Leu Glu Ile Pro Ala Val Ser His Val Cys Ala Asp Leu Leu Asp 
        115                 120                 125 

Arg Gln Ile Leu Ala Ala Asp Ala Gly Ala Asp Ala Gly Gln Leu Asp 
    130                 135                 140 

Leu Val Asp Gln Ile Asp Gln Arg Asn Leu Leu Arg Ala Lys Glu Tyr 
145                 150                 155                 160 

Leu Glu Phe Phe Gln Ser Asn Pro Met Asn Ser Leu Pro Pro Ala Ala 
                165                 170                 175 

Ala Ala Ala Ala Ala Ser Phe Pro Trp Ser Ala Phe Gly Ala Ser Asp 
            180                 185                 190 

Asp Asp Leu Asp Ala Thr Lys Glu Ala Val Ala Ala Ala Val Ala Ala 
        195                 200                 205 

Val Ala Ala Gly Asp Cys Asn Gly Leu Asp Phe Tyr Gly Pro Gly Pro 
    210                 215                 220 

Pro Ala Glu Arg Pro Pro Thr Gly Asp Gly Asp Glu Gly Asp Ser Asn 
225                 230                 235                 240 

Pro Gly Leu Trp Pro Glu Arg Asp Glu Asp Ala Pro Thr Gly Gly Leu 
                245                 250                 255 

Phe Pro Pro Pro Val Ala Pro Pro Ala Ala Thr Gln Asn Gly His Tyr 
            260                 265                 270 

Gly Arg Gly Gly Glu Glu Glu Ala Ala Ser Leu Ser Glu Ala Ala Pro 
        275                 280                 285 

Glu Pro Gly Asp Ser Pro Gly Phe Leu Ser Gly Ala Ala Glu Gly Glu 
    290                 295                 300 

Asp Gly Asp Gly Pro Asp Val Asp Gly Leu Ala Ala Ser Thr Leu Leu 
305                 310                 315                 320 

Gln Gln Met Met Ser Ser Val Gly Arg Ala Gly Ala Ala Ala Gly Asp 
                325                 330                 335 

Ser Asp Glu Glu Ser Arg Ala Asp Asp Lys Gly Val Met Asp Tyr Tyr 
            340                 345                 350 

Leu Lys Tyr Phe Ser Gly Ala His Asp Gly Asp Val Tyr Pro Ala Trp 
        355                 360                 365 

Ser Gln Lys Val Glu Lys Lys Ile Arg Ala Lys Ala Phe Gln Lys Cys 
    370                 375                 380 

Pro Ile Cys Glu Lys Val Ile Gln Gly Ala Gly Lys Leu Pro Arg His 
385                 390                 395                 400 

Ile Arg Thr His Thr Gly Glu Lys Pro Tyr Glu Cys Asn Ile Cys Lys 
                405                 410                 415 

Val Arg Phe Thr Arg Gln Asp Lys Leu Lys Val His Met Arg Lys His 
            420                 425                 430 

Thr Gly Glu Lys Pro Tyr Leu Cys Gln Gln Cys Gly Ala Ala Phe Ala 
        435                 440                 445 

His Asn Tyr Asp Leu Lys Asn His Met Arg Val His Thr Gly Leu Arg 
    450                 455                 460 

Pro Tyr Gln Cys Asp Ser Cys Cys Lys Thr Phe Val Arg Ser Asp His 
465                 470                 475                 480 

Leu His Arg His Leu Lys Lys Asp Gly Cys Asn Gly Val Pro Ser Arg 
                485                 490                 495 

Arg Gly Arg Lys Pro Arg Val Arg Gly Gly Ala Pro Asp Pro Ser Pro 
            500                 505                 510 

Gly Ala Thr Ala Thr Pro Gly Ala Pro Ala Gln Pro Ser Ser Pro Asp 
        515                 520                 525 

Ala Arg Arg Asn Gly Gln Glu Lys His Phe Lys Asp Glu Asp Glu Asp 
    530                 535                 540 

Glu Asp Val Ala Ser Pro Asp Gly Leu Gly Arg Leu Asn Val Ala Gly 
545                 550                 555                 560 

Ala Gly Gly Gly Gly Asp Ser Gly Gly Gly Pro Gly Ala Ala Thr Asp 
                565                 570                 575 

Gly Asn Phe Thr Ala Gly Leu Ala Glu Phe Met Glu Pro Val Asn Pro 
            580                 585                 590 

Ser Leu Glu Pro Trp Lys His Pro Gly Ser Gln Pro Lys Thr Ala Cys 
        595                 600                 605 

Thr Asn Cys Tyr Cys Ala Lys Cys Cys Phe His Cys Gln Val Cys Phe 
    610                 615                 620 

Ile Thr Lys Ala Leu Gly Ile Ser Tyr Gly Arg Ala Lys Arg Arg Gln 
625                 630                 635                 640 

Arg Arg Arg Pro Pro Gln Gly Ser Gln Thr His Gln Val Ser Leu Ser 
                645                 650                 655 

Lys Leu Ile 

 
           
             7  
             560  
             PRT  
             Artificial Sequence  
             
               TAT dMt-MeCP2 fusion protein  
             
           
            7 

Met Glu Pro Val Asn Pro Ser Leu Glu Pro Trp Lys His Pro Gly Ser 
  1               5                  10                  15 

Gln Pro Lys Thr Ala Cys Thr Asn Cys Tyr Cys Ala Lys Cys Cys Phe 
             20                  25                  30 

His Cys Gln Val Cys Phe Ile Thr Lys Ala Leu Gly Ile Ser Tyr Gly 
         35                  40                  45 

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

His Gln Val Ser Leu Ser Lys Pro Glu Phe Met Val Ala Gly Met Leu 
 65                  70                  75                  80 

Gly Leu Arg Glu Glu Lys Ser Glu Asp Gln Asp Leu Gln Gly Leu Lys 
                 85                  90                  95 

Asp Lys Pro Leu Lys Phe Lys Lys Val Lys Lys Asp Lys Lys Glu Glu 
            100                 105                 110 

Lys Glu Gly Lys His Glu Pro Val Gln Pro Ser Ala His His Ser Ala 
        115                 120                 125 

Glu Pro Ala Glu Ala Gly Lys Ala Glu Thr Ser Glu Gly Ser Gly Ser 
    130                 135                 140 

Ala Pro Ala Val Pro Glu Ala Ser Ala Ser Pro Lys Gln Arg Arg Ser 
145                 150                 155                 160 

Ile Ile Arg Asp Arg Gly Pro Met Tyr Asp Asp Pro Thr Leu Pro Glu 
                165                 170                 175 

Gly Trp Thr Arg Lys Leu Lys Gln Arg Lys Ser Gly Arg Ser Ala Gly 
            180                 185                 190 

Lys Tyr Asp Val Tyr Leu Ile Asn Pro Gln Gly Lys Ala Phe Arg Ser 
        195                 200                 205 

Lys Val Glu Leu Ile Ala Tyr Phe Glu Lys Val Gly Asp Thr Ser Leu 
    210                 215                 220 

Asp Pro Asn Asp Phe Asp Phe Thr Val Thr Gly Arg Gly Ser Pro Ser 
225                 230                 235                 240 

Arg Arg Glu Gln Lys Pro Pro Lys Lys Pro Lys Ser Pro Lys Ala Pro 
                245                 250                 255 

Gly Thr Gly Arg Gly Arg Gly Arg Pro Lys Gly Ser Gly Thr Thr Arg 
            260                 265                 270 

Pro Lys Ala Ala Thr Ser Glu Gly Val Gln Val Lys Arg Val Leu Glu 
        275                 280                 285 

Lys Ser Pro Gly Lys Leu Leu Val Lys Met Pro Phe Gln Thr Ser Pro 
    290                 295                 300 

Gly Gly Lys Ala Glu Gly Gly Gly Ala Thr Thr Ser Thr Gln Val Met 
305                 310                 315                 320 

Val Ile Lys Arg Pro Gly Arg Lys Arg Lys Ala Glu Ala Asp Pro Gln 
                325                 330                 335 

Ala Ile Pro Lys Lys Arg Gly Arg Lys Pro Gly Ser Val Val Ala Ala 
            340                 345                 350 

Ala Ala Ala Glu Ala Lys Lys Lys Ala Val Lys Glu Ser Ser Ile Arg 
        355                 360                 365 

Ser Val Gln Glu Thr Val Leu Pro Ile Lys Lys Arg Lys Thr Arg Glu 
    370                 375                 380 

Thr Val Ser Ile Glu Val Lys Glu Val Val Lys Pro Leu Leu Val Ser 
385                 390                 395                 400 

Thr Leu Gly Glu Lys Ser Gly Lys Gly Leu Lys Thr Cys Lys Ser Pro 
                405                 410                 415 

Gly Arg Lys Ser Lys Glu Ser Ser Pro Lys Gly Arg Ser Ser Ser Ala 
            420                 425                 430 

Ser Ser Pro Pro Lys Lys Glu His His His His His His His Ser Glu 
        435                 440                 445 

Ser Pro Lys Ala Pro Val Pro Leu Leu Pro Pro Leu Pro Pro Pro Pro 
    450                 455                 460 

Pro Glu Pro Glu Ser Ser Glu Asp Pro Thr Ser Pro Pro Glu Pro Gln 
465                 470                 475                 480 

Asp Leu Ser Ser Ser Val Cys Lys Glu Glu Lys Met Pro Arg Gly Gly 
                485                 490                 495 

Ser Leu Glu Ser Asp Gly Cys Pro Lys Glu Pro Ala Lys Thr Gln Pro 
            500                 505                 510 

Ala Val Ala Thr Ala Ala Thr Ala Ala Glu Lys Tyr Lys His Arg Gly 
        515                 520                 525 

Glu Gly Glu Arg Lys Asp Ile Val Ser Ser Ser Met Pro Arg Pro Asn 
    530                 535                 540 

Arg Glu Glu Pro Val Asp Ser Arg Thr Pro Val Thr Glu Arg Val Ser 
545                 550                 555                 560 

 
           
             8  
             556  
             PRT  
             Artificial Sequence  
             
               TAT dMt-HDAC1 fusion protein  
             
           
            8 

Met Glu Pro Val Asn Pro Ser Leu Glu Pro Trp Lys His Pro Gly Ser 
  1               5                  10                  15 

Gln Pro Lys Thr Ala Cys Thr Asn Cys Tyr Cys Ala Lys Cys Cys Phe 
             20                  25                  30 

His Cys Gln Val Cys Phe Ile Thr Lys Ala Leu Gly Ile Ser Tyr Gly 
         35                  40                  45 

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

His Gln Val Ser Leu Ser Lys Pro Glu Phe Met Ala Gln Thr Gln Gly 
 65                  70                  75                  80 

Thr Arg Arg Lys Val Cys Tyr Tyr Tyr Asp Gly Asp Val Gly Asn Tyr 
                 85                  90                  95 

Tyr Tyr Gly Gln Gly His Pro Met Lys Pro His Arg Ile Arg Met Thr 
            100                 105                 110 

His Asn Leu Leu Leu Asn Tyr Gly Leu Tyr Arg Lys Met Glu Ile Tyr 
        115                 120                 125 

Arg Pro His Lys Ala Asn Ala Glu Glu Met Thr Lys Tyr His Ser Asp 
    130                 135                 140 

Asp Tyr Ile Lys Phe Leu Arg Ser Ile Arg Pro Asp Asn Met Ser Glu 
145                 150                 155                 160 

Tyr Ser Lys Gln Met Gln Arg Phe Asn Val Gly Glu Asp Cys Pro Val 
                165                 170                 175 

Phe Asp Gly Leu Phe Glu Phe Cys Gln Leu Ser Thr Gly Gly Ser Val 
            180                 185                 190 

Ala Ser Ala Val Lys Leu Asn Lys Gln Gln Thr Asp Ile Ala Val Asn 
        195                 200                 205 

Trp Ala Gly Gly Leu His His Ala Lys Lys Ser Glu Ala Ser Gly Phe 
    210                 215                 220 

Cys Tyr Val Asn Asp Ile Val Leu Ala Ile Leu Glu Leu Leu Lys Tyr 
225                 230                 235                 240 

His Gln Arg Val Leu Tyr Ile Asp Ile Asp Ile His His Gly Asp Gly 
                245                 250                 255 

Val Glu Glu Ala Phe Tyr Thr Thr Asp Arg Val Met Thr Val Ser Phe 
            260                 265                 270 

His Lys Tyr Gly Glu Tyr Phe Pro Gly Thr Gly Asp Leu Arg Asp Ile 
        275                 280                 285 

Gly Ala Gly Lys Gly Lys Tyr Tyr Ala Val Asn Tyr Pro Leu Arg Asp 
    290                 295                 300 

Gly Ile Asp Asp Glu Ser Tyr Glu Ala Ile Phe Lys Pro Val Met Ser 
305                 310                 315                 320 

Lys Val Met Glu Met Phe Gln Pro Ser Ala Val Val Leu Gln Cys Gly 
                325                 330                 335 

Ser Asp Ser Leu Ser Gly Asp Arg Leu Gly Cys Phe Asn Leu Thr Ile 
            340                 345                 350 

Lys Gly His Ala Lys Cys Val Glu Phe Val Lys Ser Phe Asn Leu Pro 
        355                 360                 365 

Met Leu Met Leu Gly Gly Gly Gly Tyr Thr Ile Arg Asn Val Ala Arg 
    370                 375                 380 

Cys Trp Thr Tyr Glu Thr Ala Val Ala Leu Asp Thr Glu Ile Pro Asn 
385                 390                 395                 400 

Glu Leu Pro Tyr Asn Asp Tyr Phe Glu Tyr Phe Gly Pro Asp Phe Lys 
                405                 410                 415 

Leu His Ile Ser Pro Ser Asn Met Thr Asn Gln Asn Thr Asn Glu Tyr 
            420                 425                 430 

Leu Glu Lys Ile Lys Gln Arg Leu Phe Glu Asn Leu Arg Met Leu Pro 
        435                 440                 445 

His Ala Pro Gly Val Gln Met Gln Ala Ile Pro Glu Asp Ala Ile Pro 
    450                 455                 460 

Glu Glu Ser Gly Asp Glu Asp Glu Asp Asp Pro Asp Lys Arg Ile Ser 
465                 470                 475                 480 

Ile Cys Ser Ser Asp Lys Arg Ile Ala Cys Glu Glu Glu Phe Ser Asp 
                485                 490                 495 

Ser Glu Glu Glu Gly Glu Gly Gly Arg Lys Asn Ser Ser Asn Phe Lys 
            500                 505                 510 

Lys Ala Lys Arg Val Lys Thr Glu Asp Glu Lys Glu Lys Asp Pro Glu 
        515                 520                 525 

Glu Lys Lys Glu Val Thr Glu Glu Glu Lys Thr Lys Glu Glu Lys Pro 
    530                 535                 540 

Glu Ala Lys Gly Val Lys Glu Glu Val Lys Leu Ala 
545                 550                 555 

 
           
             9  
             204  
             PRT  
             Artificial Sequence  
             
               TAT dMt-POZ fusion protein  
             
           
            9 

Met Glu Pro Val Asn Pro Ser Leu Glu Pro Trp Lys His Pro Gly Ser 
  1               5                  10                  15 

Gln Pro Lys Thr Ala Cys Thr Asn Cys Tyr Cys Ala Lys Cys Cys Phe 
             20                  25                  30 

His Cys Gln Val Cys Phe Ile Thr Lys Ala Leu Gly Ile Ser Tyr Gly 
         35                  40                  45 

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

His Gln Val Ser Leu Ser Lys Pro Glu Phe Met Ala Gly Gly Val Asp 
 65                  70                  75                  80 

Gly Pro Ile Gly Ile Pro Phe Pro Asp His Ser Ser Asp Ile Leu Ser 
                 85                  90                  95 

Gly Leu Asn Glu Gln Arg Thr Gln Gly Leu Leu Cys Asp Val Val Ile 
            100                 105                 110 

Leu Val Glu Gly Arg Glu Phe Pro Thr His Arg Ser Val Leu Ala Ala 
        115                 120                 125 

Cys Ser Gln Tyr Phe Lys Lys Leu Phe Thr Ser Gly Ala Val Val Asp 
    130                 135                 140 

Gln Gln Asn Val Tyr Glu Ile Asp Phe Val Ser Ala Glu Ala Leu Thr 
145                 150                 155                 160 

Ala Leu Met Asp Phe Ala Tyr Thr Ala Thr Leu Thr Val Ser Thr Ala 
                165                 170                 175 

Asn Val Gly Asp Ile Leu Ser Ala Ala Arg Leu Leu Glu Ile Pro Ala 
            180                 185                 190 

Val Ser His Val Cys Ala Asp Leu Leu Asp Arg Gln 
        195                 200 

 
           
             10  
             658  
             PRT  
             Artificial Sequence  
             
               TAT dMt-FBI-1 fusion protein  
             
           
            10 

Met Glu Pro Val Asn Pro Ser Leu Glu Pro Trp Lys His Pro Gly Ser 
  1               5                  10                  15 

Gln Pro Lys Thr Ala Cys Thr Asn Cys Tyr Cys Ala Lys Cys Cys Phe 
             20                  25                  30 

His Cys Gln Val Cys Phe Ile Thr Lys Ala Leu Gly Ile Ser Tyr Gly 
         35                  40                  45 

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

His Gln Val Ser Leu Ser Lys Pro Glu Phe Met Ala Gly Gly Val Asp 
 65                  70                  75                  80 

Gly Pro Ile Gly Ile Pro Phe Pro Asp His Ser Ser Asp Ile Leu Ser 
                 85                  90                  95 

Gly Leu Asn Glu Gln Arg Thr Gln Gly Leu Leu Cys Asp Val Val Ile 
            100                 105                 110 

Leu Val Glu Gly Arg Glu Phe Pro Thr His Arg Ser Val Leu Ala Ala 
        115                 120                 125 

Cys Ser Gln Tyr Phe Lys Lys Leu Phe Thr Ser Gly Ala Val Val Asp 
    130                 135                 140 

Gln Gln Asn Val Tyr Glu Ile Asp Phe Val Ser Ala Glu Ala Leu Thr 
145                 150                 155                 160 

Ala Leu Met Asp Phe Ala Tyr Thr Ala Thr Leu Thr Val Ser Thr Ala 
                165                 170                 175 

Asn Val Gly Asp Ile Leu Ser Ala Ala Arg Leu Leu Glu Ile Pro Ala 
            180                 185                 190 

Val Ser His Val Cys Ala Asp Leu Leu Asp Arg Gln Ile Leu Ala Ala 
        195                 200                 205 

Asp Ala Gly Ala Asp Ala Gly Gln Leu Asp Leu Val Asp Gln Ile Asp 
    210                 215                 220 

Gln Arg Asn Leu Leu Arg Ala Lys Glu Tyr Leu Glu Phe Phe Gln Ser 
225                 230                 235                 240 

Asn Pro Met Asn Ser Leu Pro Pro Ala Ala Ala Ala Ala Ala Ala Ser 
                245                 250                 255 

Phe Pro Trp Ser Ala Phe Gly Ala Ser Asp Asp Asp Leu Asp Ala Thr 
            260                 265                 270 

Lys Glu Ala Val Ala Ala Ala Val Ala Ala Val Ala Ala Gly Asp Cys 
        275                 280                 285 

Asn Gly Leu Asp Phe Tyr Gly Pro Gly Pro Pro Ala Glu Arg Pro Pro 
    290                 295                 300 

Thr Gly Asp Gly Asp Glu Gly Asp Ser Asn Pro Gly Leu Trp Pro Glu 
305                 310                 315                 320 

Arg Asp Glu Asp Ala Pro Thr Gly Gly Leu Phe Pro Pro Pro Val Ala 
                325                 330                 335 

Pro Pro Ala Ala Thr Gln Asn Gly His Tyr Gly Arg Gly Gly Glu Glu 
            340                 345                 350 

Glu Ala Ala Ser Leu Ser Glu Ala Ala Pro Glu Pro Gly Asp Ser Pro 
        355                 360                 365 

Gly Phe Leu Ser Gly Ala Ala Glu Gly Glu Asp Gly Asp Gly Pro Asp 
    370                 375                 380 

Val Asp Gly Leu Ala Ala Ser Thr Leu Leu Gln Gln Met Met Ser Ser 
385                 390                 395                 400 

Val Gly Arg Ala Gly Ala Ala Ala Gly Asp Ser Asp Glu Glu Ser Arg 
                405                 410                 415 

Ala Asp Asp Lys Gly Val Met Asp Tyr Tyr Leu Lys Tyr Phe Ser Gly 
            420                 425                 430 

Ala His Asp Gly Asp Val Tyr Pro Ala Trp Ser Gln Lys Val Glu Lys 
        435                 440                 445 

Lys Ile Arg Ala Lys Ala Phe Gln Lys Cys Pro Ile Cys Glu Lys Val 
    450                 455                 460 

Ile Gln Gly Ala Gly Lys Leu Pro Arg His Ile Arg Thr His Thr Gly 
465                 470                 475                 480 

Glu Lys Pro Tyr Glu Cys Asn Ile Cys Lys Val Arg Phe Thr Arg Gln 
                485                 490                 495 

Asp Lys Leu Lys Val His Met Arg Lys His Thr Gly Glu Lys Pro Tyr 
            500                 505                 510 

Leu Cys Gln Gln Cys Gly Ala Ala Phe Ala His Asn Tyr Asp Leu Lys 
        515                 520                 525 

Asn His Met Arg Val His Thr Gly Leu Arg Pro Tyr Gln Cys Asp Ser 
    530                 535                 540 

Cys Cys Lys Thr Phe Val Arg Ser Asp His Leu His Arg His Leu Lys 
545                 550                 555                 560 

Lys Asp Gly Cys Asn Gly Val Pro Ser Arg Arg Gly Arg Lys Pro Arg 
                565                 570                 575 

Val Arg Gly Gly Ala Pro Asp Pro Ser Pro Gly Ala Thr Ala Thr Pro 
            580                 585                 590 

Gly Ala Pro Ala Gln Pro Ser Ser Pro Asp Ala Arg Arg Asn Gly Gln 
        595                 600                 605 

Glu Lys His Phe Lys Asp Glu Asp Glu Asp Glu Asp Val Ala Ser Pro 
    610                 615                 620 

Asp Gly Leu Gly Arg Leu Asn Val Ala Gly Ala Gly Gly Gly Gly Asp 
625                 630                 635                 640 

Ser Gly Gly Gly Pro Gly Ala Ala Thr Asp Gly Asn Phe Thr Ala Gly 
                645                 650                 655 

Leu Ala 

 
           
             11  
             1686  
             DNA  
             Artificial Sequence  
             
               base sequence of MeCP2-TAT dMt fusion protein  
             
           
            11 

atggtagctg ggatgttagg gctcagggaa gaaaagtcag aagaccagga cctccagggc     60 

ctcaaggaca aacccctcaa gtttaaaaag gtgaagaaag ataagaaaga agagaaagag    120 

ggcaagcatg agcccgtgca gccatcagcc caccactctg ctgagcccgc agaggcaggc    180 

aaagcagaga catcagaagg gtcaggctcc gccccggctg tgccggaagc ttctgcctcc    240 

cccaaacagc ggcgctccat catccgtgac cggggaccca tgtatgatga ccccaccctg    300 

cctgaaggct ggacacggaa gcttaagcaa aggaaatctg gccgctctgc tgggaagtat    360 

gatgtgtatt tgatcaatcc ccagggaaaa gcctttcgct ctaaagtgga gttgattgcg    420 

tacttcgaaa aggtaggcga cacatccctg gaccctaatg attttgactt cacggtaact    480 

gggagaggga gcccctcccg gcgagagcag aaaccaccta agaagcccaa atctcccaaa    540 

gctccaggaa ctggcagagg ccggggacgc cccaaaggga gcggcaccac gagacccaag    600 

gcggccacgt cagagggtgt gcaggtgaaa agggtcctgg agaaaagtcc tgggaagctc    660 

cttgtcaaga tgccttttca aacttcgcca gggggcaagg ctgagggggg tggggccacc    720 

acatccaccc aggtcatggt gatcaaacgc cccggcagga agcgaaaagc tgaagctgac    780 

cctcaggcca ttcccaagaa acggggccga aagccgggga gtgtggtggc agccgctgcc    840 

gccgaggcca aaaagaaagc cgtgaaggag tcttctatcc gatctgtgca ggagaccgta    900 

ctccccatca agaagcgcaa gacccgggag acggtcagca tcgaggtcaa ggaagtggtg    960 

aagcccctgc tggtgtccac cctcggtgag aagagcggga aaggactgaa gacctgtaag   1020 

agccctgggc ggaaaagcaa ggagagcagc cccaaggggc gcagcagcag cgcctcctca   1080 

ccccccaaga aggagcacca ccaccatcac caccactcag agtccccaaa ggcccccgtg   1140 

ccactgctcc cacccctgcc cccacctcca cctgagcccg agagctccga ggaccccacc   1200 

agcccccctg agccccagga cttgagcagc agcgtctgca aagaggagaa gatgcccaga   1260 

ggaggctcac tggagagcga cggctgcccc aaggagccag ctaagactca gcccgcggtt   1320 

gccaccgccg ccacggccgc agaaaagtac aaacaccgag gggagggaga gcgcaaagac   1380 

attgtttcat cctccatgcc aaggccaaac agagaggagc ctgtggacag ccggacgccc   1440 

gtgaccgaga gagttagcga attcatggag ccagtaaatc ctagcctaga gccctggaag   1500 

catccaggaa gtcagcctaa aactgcttgt accaattgct attgtgcaaa gtgttgcttt   1560 

cattgccaag tttgtttcat aacaaaagcc ttaggcatct cctatggcag ggcaaagcgg   1620 

agacagcgac gaagacctcc tcaaggcagt cagactcatc aagtttctct atcaaagctg   1680 

atctag                                                              1686 

 
           
             12  
             1674  
             DNA  
             Artificial Sequence  
             
               base sequence of HDAC1-TAT dMt fusion protein  
             
           
            12 

atggcgcaga cgcagggcac ccggaggaaa gtctgttact actacgacgg ggatgttgga     60 

aattactatt atggacaagg ccacccaatg aagcctcacc gaatccgcat gactcataat    120 

ttgctgctca actatggtct ctaccgaaaa atggaaatct atcgccctca caaagccaat    180 

gctgaggaga tgaccaagta ccacagcgat gactacatta aattcttgcg ctccatccgt    240 

ccagataaca tgtcggagta cagcaagcag atgcagagat tcaacgttgg tgaggactgt    300 

ccagtattcg atggcctgtt tgagttctgt cagttgtcta ctggtggttc tgtggcaagt    360 

gctgtgaaac ttaataagca gcagacggac atcgctgtga attgggctgg gggcctgcac    420 

catgcaaaga agtccgaggc atctggcttc tgttacgtca atgatatcgt cttggccatc    480 

ctggaactgc taaagtatca ccagagggtg ctgtacattg acattgatat tcaccatggt    540 

gacggcgtgg aagaggcctt ctacaccacg gaccgggtca tgactgtgtc ctttcataag    600 

tatggagagt acttcccagg aactggggac ctacgggata tcggggctgg caaaggcaag    660 

tattatgctg ttaactaccc gctccgagac gggattgatg acgagtccta tgaggccatt    720 

ttcaagccgg tcatgtccaa agtaatggag atgttccagc ctagtgcggt ggtcttacag    780 

tgtggctcag actccctatc tggggatcgg ttaggttgct tcaatctaac tatcaaagga    840 

cacgccaagt gtgtggaatt tgtcaagagc tttaacctgc ctatgctgat gctgggaggc    900 

ggtggttaca ccattcgtaa cgttgcccgg tgctggacat atgagacagc tgtggccctg    960 

gatacggaga tccctaatga gcttccatac aatgactact ttgaatactt tggaccagat   1020 

ttcaagctcc acatcagtcc ttccaatatg actaaccaga acacgaatga gtacctggag   1080 

aagatcaaac agcgactgtt tgagaacctt agaatgctgc cgcacgcacc tggggtccaa   1140 

atgcaggcga ttcctgagga cgccatccct gaggagagtg gcgatgagga cgaagacgac   1200 

cctgacaagc gcatctcgat ctgctcctct gacaaacgaa ttgcctgtga ggaagagttc   1260 

tccgattctg aagaggaggg agaggggggc cgcaagaact cttccaactt caaaaaagcc   1320 

aagagagtca aaacagagga tgaaaaagag aaagacccag aggagaagaa agaagtcacc   1380 

gaagaggaga aaaccaagga ggagaagcca gaagccaaag gggtcaagga ggaggtcaag   1440 

ttggccgaat tcatggagcc agtaaatcct agcctagagc cctggaagca tccaggaagt   1500 

cagcctaaaa ctgcttgtac caattgctat tgtgcaaagt gttgctttca ttgccaagtt   1560 

tgtttcataa caaaagcctt aggcatctcc tatggcaggg caaagcggag acagcgacga   1620 

agacctcctc aaggcagtca gactcatcaa gtttctctat caaagctgat ctag         1674 

 
           
             13  
             618  
             DNA  
             Artificial Sequence  
             
               base sequence of POZ-TAT dMt fusion protein  
             
           
            13 

atggccggcg gcgtggacgg ccccatcggg atcccgttcc ccgaccacag cagcgacatc     60 

ctgagtgggc tgaacgagca gcggacgcag ggcctgctgt gcgacgtggt gatcctggtg    120 

gagggccgcg agttccccac gcaccgctcg gtgctggccg cctgcagcca gtacttcaag    180 

aagctgttca cgtcgggcgc cgtggtggac cagcagaacg tgtacgagat cgacttcgtc    240 

agcgccgagg cgctcaccgc gctcatggac ttcgcctaca cggccacgct caccgtcagc    300 

acagccaacg tgggtgacat cctcagcgcc gcccgcctgc tggagatccc cgccgtgagc    360 

cacgtgtgcg ccgacctcct ggaccggcag gaattcatgg agccagtaaa tcctagccta    420 

gagccctgga agcatccagg aagtcagcct aaaactgctt gtaccaattg ctattgtgca    480 

aagtgttgct ttcattgcca agtttgtttc ataacaaaag ccttaggcat ctcctatggc    540 

agggcaaagc ggagacagcg acgaagacct cctcaaggca gtcagactca tcaagtttct    600 

ctatcaaagc tgatctag                                                  618 

 
           
             14  
             1980  
             DNA  
             Artificial Sequence  
             
               base sequence of FBI-1-TAT dMt fusion protein  
             
           
            14 

atggccggcg gcgtggacgg ccccatcggg atcccgttcc ccgaccacag cagcgacatc     60 

ctgagtgggc tgaacgagca gcggacgcag ggcctgctgt gcgacgtggt gatcctggtg    120 

gagggccgcg agttccccac gcaccgctcg gtgctggccg cctgcagcca gtacttcaag    180 

aagctgttca cgtcgggcgc cgtggtggac cagcagaacg tgtacgagat cgacttcgtc    240 

agcgccgagg cgctcaccgc gctcatggac ttcgcctaca cggccacgct caccgtcagc    300 

acagccaacg tgggtgacat cctcagcgcc gcccgcctgc tggagatccc cgccgtgagc    360 

cacgtgtgcg ccgacctcct ggaccggcag atcctggcgg ccgacgcggg cgccgacgcc    420 

gggcagctgg accttgtaga tcaaattgat cagcgcaacc tcctccgcgc caaggagtac    480 

ctcgagttct tccagagcaa ccccatgaac agcctgcccc ccgcggccgc cgccgccgct    540 

gccagcttcc cgtggtccgc ctttggggcg tccgatgatg acctggatgc caccaaggag    600 

gccgtggccg ccgctgtggc cgccgtggcc gcgggcgact gcaacggctt agacttctat    660 

gggccgggcc ccccggccga gcggcccccg acgggggacg gggacgaggg cgacagcaac    720 

ccgggtctgt ggccagagcg ggatgaggac gcccccaccg ggggtctctt tccgccgccg    780 

gtggccccgc cggccgccac gcagaacggc cactacggcc gcggcggaga ggaggaggcc    840 

gcctcgctgt cggaggcggc ccccgagccg ggcgactctc cgggcttcct gtcgggagcg    900 

gccgagggcg aggacgggga cgggcccgac gtggacgggc tggcggccag cacgctgctg    960 

cagcagatga tgtcatcggt gggccgggcg ggggccgcgg cgggggacag cgacgaggag   1020 

tcgcgggccg acgacaaggg cgtcatggac tactacctga agtacttcag cggcgcccac   1080 

gacggcgacg tctacccggc ctggtcgcag aaggtggaga agaagatccg agccaaggcc   1140 

ttccagaagt gccccatctg cgagaaggtc atccagggcg ccggcaagct gccgcgacac   1200 

atccgcaccc acacgggcga gaagccctac gagtgcaaca tctgcaaggt ccgcttcacc   1260 

aggcaggaca agctgaaggt gcacatgcgg aagcacacgg gcgagaagcc gtacctgtgc   1320 

cagcagtgcg gcgccgcctt tgcccacaac tacgacctga agaaccacat gcgcgtgcac   1380 

acgggcctgc gcccctacca gtgcgacagc tgctgcaaga ccttcgtccg ctccgaccac   1440 

ctgcacagac acctcaagaa agacggctgc aacggcgtcc cctcgcgccg cggccgcaag   1500 

ccccgcgtcc ggggcggggc gcccgacccc agcccggggg ccaccgcgac ccccggcgcc   1560 

cccgcccagc ccagctcccc cgacgcccgg cgcaacggcc aggagaagca ctttaaggac   1620 

gaggacgagg acgaggacgt ggccagcccc gacggcttgg gccggttgaa tgtagcgggc   1680 

gccggtggag gaggtgacag cggaggtggc cccggggccg ccaccgacgg taacttcaca   1740 

gccggactcg ccgaattcat ggagccagta aatcctagcc tagagccctg gaagcatcca   1800 

ggaagtcagc ctaaaactgc ttgtaccaat tgctattgtg caaagtgttg ctttcattgc   1860 

caagtttgtt tcataacaaa agccttaggc atctcctatg gcagggcaaa gcggagacag   1920 

cgacgaagac ctcctcaagg cagtcagact catcaagttt ctctatcaaa gctgatctag   1980 

                                                                    1980 

 
           
             15  
             1683  
             DNA  
             Artificial Sequence  
             
               base sequence of TAT dMt-MeCP2 fusion protein  
             
           
            15 

atggagccag taaatcctag cctagagccc tggaagcatc caggaagtca gcctaaaact     60 

gcttgtacca attgctattg tgcaaagtgt tgctttcatt gccaagtttg tttcataaca    120 

aaagccttag gcatctccta tggcagggca aagcggagac agcgacgaag acctcctcaa    180 

ggcagtcaga ctcatcaagt ttctctatca aagcccgaat tcatggtagc tgggatgtta    240 

gggctcaggg aagaaaagtc agaagaccag gacctccagg gcctcaagga caaacccctc    300 

aagtttaaaa aggtgaagaa agataagaaa gaagagaaag agggcaagca tgagcccgtg    360 

cagccatcag cccaccactc tgctgagccc gcagaggcag gcaaagcaga gacatcagaa    420 

gggtcaggct ccgccccggc tgtgccggaa gcttctgcct cccccaaaca gcggcgctcc    480 

atcatccgtg accggggacc catgtatgat gaccccaccc tgcctgaagg ctggacacgg    540 

aagcttaagc aaaggaaatc tggccgctct gctgggaagt atgatgtgta tttgatcaat    600 

ccccagggaa aagcctttcg ctctaaagtg gagttgattg cgtacttcga aaaggtaggc    660 

gacacatccc tggaccctaa tgattttgac ttcacggtaa ctgggagagg gagcccctcc    720 

cggcgagagc agaaaccacc taagaagccc aaatctccca aagctccagg aactggcaga    780 

ggccggggac gccccaaagg gagcggcacc acgagaccca aggcggccac gtcagagggt    840 

gtgcaggtga aaagggtcct ggagaaaagt cctgggaagc tccttgtcaa gatgcctttt    900 

caaacttcgc cagggggcaa ggctgagggg ggtggggcca ccacatccac ccaggtcatg    960 

gtgatcaaac gccccggcag gaagcgaaaa gctgaagctg accctcaggc cattcccaag   1020 

aaacggggcc gaaagccggg gagtgtggtg gcagccgctg ccgccgaggc caaaaagaaa   1080 

gccgtgaagg agtcttctat ccgatctgtg caggagaccg tactccccat caagaagcgc   1140 

aagacccggg agacggtcag catcgaggtc aaggaagtgg tgaagcccct gctggtgtcc   1200 

accctcggtg agaagagcgg gaaaggactg aagacctgta agagccctgg gcggaaaagc   1260 

aaggagagca gccccaaggg gcgcagcagc agcgcctcct caccccccaa gaaggagcac   1320 

caccaccatc accaccactc agagtcccca aaggcccccg tgccactgct cccacccctg   1380 

cccccacctc cacctgagcc cgagagctcc gaggacccca ccagcccccc tgagccccag   1440 

gacttgagca gcagcgtctg caaagaggag aagatgccca gaggaggctc actggagagc   1500 

gacggctgcc ccaaggagcc agctaagact cagcccgcgg ttgccaccgc cgccacggcc   1560 

gcagaaaagt acaaacaccg aggggaggga gagcgcaaag acattgtttc atcctccatg   1620 

ccaaggccaa acagagagga gcctgtggac agccggacgc ccgtgaccga gagagttagc   1680 

tga                                                                 1683 

 
           
             16  
             1671  
             DNA  
             Artificial Sequence  
             
               base sequence of TAT dMt-HDAC1 fusion protein  
             
           
            16 

atggagccag taaatcctag cctagagccc tggaagcatc caggaagtca gcctaaaact     60 

gcttgtacca attgctattg tgcaaagtgt tgctttcatt gccaagtttg tttcataaca    120 

aaagccttag gcatctccta tggcagggca aagcggagac agcgacgaag acctcctcaa    180 

ggcagtcaga ctcatcaagt ttctctatca aagcccgaat tcatggcgca gacgcagggc    240 

acccggagga aagtctgtta ctactacgac ggggatgttg gaaattacta ttatggacaa    300 

ggccacccaa tgaagcctca ccgaatccgc atgactcata atttgctgct caactatggt    360 

ctctaccgaa aaatggaaat ctatcgccct cacaaagcca atgctgagga gatgaccaag    420 

taccacagcg atgactacat taaattcttg cgctccatcc gtccagataa catgtcggag    480 

tacagcaagc agatgcagag attcaacgtt ggtgaggact gtccagtatt cgatggcctg    540 

tttgagttct gtcagttgtc tactggtggt tctgtggcaa gtgctgtgaa acttaataag    600 

cagcagacgg acatcgctgt gaattgggct gggggcctgc accatgcaaa gaagtccgag    660 

gcatctggct tctgttacgt caatgatatc gtcttggcca tcctggaact gctaaagtat    720 

caccagaggg tgctgtacat tgacattgat attcaccatg gtgacggcgt ggaagaggcc    780 

ttctacacca cggaccgggt catgactgtg tcctttcata agtatggaga gtacttccca    840 

ggaactgggg acctacggga tatcggggct ggcaaaggca agtattatgc tgttaactac    900 

ccgctccgag acgggattga tgacgagtcc tatgaggcca ttttcaagcc ggtcatgtcc    960 

aaagtaatgg agatgttcca gcctagtgcg gtggtcttac agtgtggctc agactcccta   1020 

tctggggatc ggttaggttg cttcaatcta actatcaaag gacacgccaa gtgtgtggaa   1080 

tttgtcaaga gctttaacct gcctatgctg atgctgggag gcggtggtta caccattcgt   1140 

aacgttgccc ggtgctggac atatgagaca gctgtggccc tggatacgga gatccctaat   1200 

gagcttccat acaatgacta ctttgaatac tttggaccag atttcaagct ccacatcagt   1260 

ccttccaata tgactaacca gaacacgaat gagtacctgg agaagatcaa acagcgactg   1320 

tttgagaacc ttagaatgct gccgcacgca cctggggtcc aaatgcaggc gattcctgag   1380 

gacgccatcc ctgaggagag tggcgatgag gacgaagacg accctgacaa gcgcatctcg   1440 

atctgctcct ctgacaaacg aattgcctgt gaggaagagt tctccgattc tgaagaggag   1500 

ggagaggggg gccgcaagaa ctcttccaac ttcaaaaaag ccaagagagt caaaacagag   1560 

gatgaaaaag agaaagaccc agaggagaag aaagaagtca ccgaagagga gaaaaccaag   1620 

gaggagaagc cagaagccaa aggggtcaag gaggaggtca agttggcctg a            1671 

 
           
             17  
             612  
             DNA  
             Artificial Sequence  
             
               base sequence of TAT dMt-POZ fusion protein  
             
           
            17 

atggagccag taaatcctag cctagagccc tggaagcatc caggaagtca gcctaaaact     60 

gcttgtacca attgctattg tgcaaagtgt tgctttcatt gccaagtttg tttcataaca    120 

aaagccttag gcatctccta tggcagggca aagcggagac agcgacgaag acctcctcaa    180 

ggcagtcaga ctcatcaagt ttctctatca aagcccgaat tcatggccgg cggcgtggac    240 

ggccccatcg ggatcccgtt ccccgaccac agcagcgaca tcctgagtgg gctgaacgag    300 

cagcggacgc agggcctgct gtgcgacgtg gtgatcctgg tggagggccg cgagttcccc    360 

acgcaccgct cggtgctggc cgcctgcagc cagtacttca agaagctgtt cacgtcgggc    420 

gccgtggtgg accagcagaa cgtgtacgag atcgacttcg tcagcgccga ggcgctcacc    480 

gcgctcatgg acttcgccta cacggccacg ctcaccgtca gcacagccaa cgtgggtgac    540 

atcctcagcg ccgcccgcct gctggagatc cccgccgtga gccacgtgtg cgccgacctc    600 

ctggaccggc ag                                                        612 

 
           
             18  
             1977  
             DNA  
             Artificial Sequence  
             
               base sequence of TAT dMt-FBI-1 fusion protein  
             
           
            18 

atggagccag taaatcctag cctagagccc tggaagcatc caggaagtca gcctaaaact     60 

gcttgtacca attgctattg tgcaaagtgt tgctttcatt gccaagtttg tttcataaca    120 

aaagccttag gcatctccta tggcagggca aagcggagac agcgacgaag acctcctcaa    180 

ggcagtcaga ctcatcaagt ttctctatca aagcccgaat tcatggccgg cggcgtggac    240 

ggccccatcg ggatcccgtt ccccgaccac agcagcgaca tcctgagtgg gctgaacgag    300 

cagcggacgc agggcctgct gtgcgacgtg gtgatcctgg tggagggccg cgagttcccc    360 

acgcaccgct cggtgctggc cgcctgcagc cagtacttca agaagctgtt cacgtcgggc    420 

gccgtggtgg accagcagaa cgtgtacgag atcgacttcg tcagcgccga ggcgctcacc    480 

gcgctcatgg acttcgccta cacggccacg ctcaccgtca gcacagccaa cgtgggtgac    540 

atcctcagcg ccgcccgcct gctggagatc cccgccgtga gccacgtgtg cgccgacctc    600 

ctggaccggc agatcctggc ggccgacgcg ggcgccgacg ccgggcagct ggaccttgta    660 

gatcaaattg atcagcgcaa cctcctccgc gccaaggagt acctcgagtt cttccagagc    720 

aaccccatga acagcctgcc ccccgcggcc gccgccgccg ctgccagctt cccgtggtcc    780 

gcctttgggg cgtccgatga tgacctggat gccaccaagg aggccgtggc cgccgctgtg    840 

gccgccgtgg ccgcgggcga ctgcaacggc ttagacttct atgggccggg ccccccggcc    900 

gagcggcccc cgacggggga cggggacgag ggcgacagca acccgggtct gtggccagag    960 

cgggatgagg acgcccccac cgggggtctc tttccgccgc cggtggcccc gccggccgcc   1020 

acgcagaacg gccactacgg ccgcggcgga gaggaggagg ccgcctcgct gtcggaggcg   1080 

gcccccgagc cgggcgactc tccgggcttc ctgtcgggag cggccgaggg cgaggacggg   1140 

gacgggcccg acgtggacgg gctggcggcc agcacgctgc tgcagcagat gatgtcatcg   1200 

gtgggccggg cgggggccgc ggcgggggac agcgacgagg agtcgcgggc cgacgacaag   1260 

ggcgtcatgg actactacct gaagtacttc agcggcgccc acgacggcga cgtctacccg   1320 

gcctggtcgc agaaggtgga gaagaagatc cgagccaagg ccttccagaa gtgccccatc   1380 

tgcgagaagg tcatccaggg cgccggcaag ctgccgcgac acatccgcac ccacacgggc   1440 

gagaagccct acgagtgcaa catctgcaag gtccgcttca ccaggcagga caagctgaag   1500 

gtgcacatgc ggaagcacac gggcgagaag ccgtacctgt gccagcagtg cggcgccgcc   1560 

tttgcccaca actacgacct gaagaaccac atgcgcgtgc acacgggcct gcgcccctac   1620 

cagtgcgaca gctgctgcaa gaccttcgtc cgctccgacc acctgcacag acacctcaag   1680 

aaagacggct gcaacggcgt cccctcgcgc cgcggccgca agccccgcgt ccggggcggg   1740 

gcgcccgacc ccagcccggg ggccaccgcg acccccggcg cccccgccca gcccagctcc   1800 

cccgacgccc ggcgcaacgg ccaggagaag cactttaagg acgaggacga ggacgaggac   1860 

gtggccagcc ccgacggctt gggccggttg aatgtagcgg gcgccggtgg aggaggtgac   1920 

agcggaggtg gccccggggc cgccaccgac ggtaacttca cagccggact cgcctaa      1977 

 
           
             19  
             37  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            19 

gatcgaattc atggagccag tacctagact agagccc                              37 

 
           
             20  
             39  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            20 

gatctctaga tcattccttc gggcctgtcg ggtcccctc                            39 

 
           
             21  
             35  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            21 

gatcgaattc atggagccag taaatcctag cctag                                35 

 
           
             22  
             35  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            22 

gatctctaga tcagctttga tagagaaact tgatg                                35 

 
           
             23  
             39  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            23 

gatcggatcc accatggtag ctgggatgtt agggctcag                            39 

 
           
             24  
             37  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            24 

gatcgaattc gctaactctc tcggtcacgg gcgtccg                              37 

 
           
             25  
             40  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            25 

gatcaagctt accatggcgc agacgcaggg cacccggagg                           40 

 
           
             26  
             41  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            26 

gatcgaattc ggccaacttg acctcctcct tgaccccttt g                         41 

 
           
             27  
             40  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            27 

gatcaagctt accatggccg gcggcgtgga cggccccatc                           40 

 
           
             28  
             38  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            28 

gatcgaattc ctgccggtcc aggaggtcgg cgcacacg                             38 

 
           
             29  
             40  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            29 

gatcaagctt accatggccg gcggcgtgga cggccccatc                           40 

 
           
             30  
             31  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            30 

gatcagattc ggcgagtccg gctgtgaagt t                                    31 

 
           
             31  
             38  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            31 

gatcggatcc accatggacg gagtaaatcc tagcctag                             38 

 
           
             32  
             34  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            32 

gatcgaattc gggctttgat agagaaactt gatg                                 34 

 
           
             33  
             35  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            33 

gatcgaattc atggtagctg ggatgttagg gctca                                35 

 
           
             34  
             35  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            34 

gatctctaga tcagctaact ctctcggtca cgggc                                35 

 
           
             35  
             35  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            35 

gatcgaattc atggcgcaga cgcagggcac ccgga                                35 

 
           
             36  
             35  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            36 

gatctctaga tcaggccaac ttgacctcct ccttg                                35 

 
           
             37  
             35  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            37 

gatcgaattc atggccggcg gcggcgtgga cggcc                                35 

 
           
             38  
             35  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            38 

gatctctaga tcactgccgg tccaggaggt cggcg                                35 

 
           
             39  
             35  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            39 

gatcgaattc atggccggcg gcggcgtgga cggcc                                35 

 
           
             40  
             34  
             DNA  
             Artificial Sequence  
             
               Primer  
             
           
            40 

gatctctaga tcaggcgagt ccggctgtga agtt                                 34