P10607119-8 A2 - Questel

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22 gtgttaattt tggtgtgatt gatgaacgat tacatcgtaa cagggaatat agagaccggt 3540 attacagaaa tctgaatata gctccggcag aggatggtta cagattagca ggtttcccac 3600 cggatcacca agcttggaga gaagaaccct ggattcatca tgcaccacaa ggttgtggaa 3660 attcatcaag aacaattaca ggtgatactt gtaatgagga gacccagaat ctgagcacaa 3720 tatatctcag gaaatatcaa tcaaaagtta agaggcagat attttcagac tatcagtcag 3780 aggttgacat atataacaga attcgggatg aattatgagg atctgggccc taacaaaaca 3840 aaaagatggg gttattccct aaacttcatg ggttacgtaa ttggaagttg ggggacattg 3900 ccacaagatc atattgtaca aaagatcaaa cactgtttta gaaaacttcc tgtaaacagg 3960 cctattgatt ggaaagtatg tcaaaggatt gtgggtcttt tgggctttgc tgctccattt 4020 acacaatgtg gatatcctgc cttaatgcct ttgtatgcat gtatacaagc taaacaggct 4080 ttcactttct cgccaactta caaggccttt ctaagtaaac agtacatgaa cctttacccc 4140 gttgctcggc aacggcctgg tctgtgccaa gtgtttgctg acgcaacccc cactggctgg 4200 ggcttggcca taggccatca gcgcatgcgt ggaacctttg tggctcctct gccgatccat 4260 actgcggaac tcctagccgc ttgttttgct cgcagccggt ctggagcaaa, gctcatagga 4320 actgacaatt ctgtcgtcct ctcgcggaaa tatacatcgt ttcgatctac gtatgatctt 4380 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc 4440 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc 4500 ggaaggaatt ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg 4560 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc 4620 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg 4680 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 4740 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 4800 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 4860 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 4920 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 4980 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 5040 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 5100 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 5160 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 5220 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 5280 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 5340 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 5400 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 5460 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 5520 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactc 62 6254 DNA Seqüência artificial Construção PJV7788 62 5578 ggcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg 60 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa 120 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc 180 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg 240 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat 300 ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca 360 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga 420 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg 480 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg 540 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata 600 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca 660 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg 720 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat 780 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga gcaagacgtt 840 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt 900 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca 960 acgtggcttt cccccccccc ccggcatgcc tgcaggtcga caatattggc tattggccat 1020 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac 1080 cgccatgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag 1140 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct 1200 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc 1260
23 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg 1320 cagtacatca agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat 1380 ggcccgcctg gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca 1440 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc 1500 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga 1560 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt 1620 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag 1680 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc 1740 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca 1800 agagtgactc accgtccgga tctcagcaag caggtatgta ctctccaggg tgggcctggc 1860 ttccccagtc aagactccag ggatttgagg gacgctgtgg gctcttctct tacatgtacc 1920 ttttgcttgc ctcaaccctg actatcttcc aggtcaggat cccagagtca ggggtctgta 1980 ttttcctgct ggtggctcca gttcaggaac agtaaaccct gctccgaata ttgcctctca 2040 catctcgtca atctccgcga ggactgggga ccctgtgacg aacatggcta gtaggtcttt 2100 gctaatcttg gtgctttgct tcctgcccct ggctgctctg ggggctagcg ctccccagtc 2160 tattacagaa ctatgttcgg aatatcgcaa cacacaaata tatacgataa atgacaagat 2220 actatcatat acggaatcga tggcaggcaa aagagaaatg gttatcatta catttaagag 2280 cggcgcaaca tttcaggtcg aagtcccggg cagtcaacat atagactccc aaaaaaaagc 2340 cattgaaagg atgaaggaca cattaagaat cacatatctg accgagacca aaattgataa 2400 attatgtgta tggaataata aaacccccaa ttcaattgcg gcaatcagta tggaaaacta 2460 gggatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga 2520 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc 2580 tctcactcgg aaggaattga gggccggccc tctgcaggtc gacaatattg gctattggcc 2640 attgcatacg ttgtatctat atcataatat gtacatttat attggctcat gtccaatatg 2700 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt 2760 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg 2820 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac 2880 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt 2940 ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca atgacggtaa 3000 atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta cttggcagta 3060 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acaccaatgg 3120 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg 3180 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata accccgcccc 3240 gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcgttt 3300 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca 3360 ccgggaccga tccagcctcc gcggccggga acggtgcatt ggaacgcgga ttccccgtgc 3420 caagagtgac tcaccgtccg gatctcagca agcaggtatg tactctccag ggtgggcctg 3480 gcttccccag tcaagactcc agggatttga gggacgctgt gggctcttct cttacatgta 3540 ccttttgctt gcctcaaccc tgactatctt ccaggtcagg atcccagagt caggggtctg 3600 tattttcctg ctggtggctc cagttcagga acagtaaacc ctgctccgaa tattgcctct 3660 cacatctcgt caatctccgc gaggactggg gaccctgtga cgaacatggc tagtaggtct 3720 ttgctaatct tggtgctttg cttcctgccc ctggctgctc tgggggctag caatggcgac 3780 aaattatacc gtgctgactc tagaccccca gatgaaataa aacgttccgg aggtcttatg 3840 cccagagggc ataatgagta cttcgataga ggaactcaaa tgaatattaa tctttatgat 3900 cacgcgagag gaacacaaac cggctttgtc agatatgatg acggatatgt ttccacttct 3960 cttagtttga gaagtgctca cttagcagga cagtctatat tatcaggata ttccacttac 4020 tatatatatg ttatagcgac agcaccaaat atgtttaatg ttaatgatgt attaggcgta 4080 tacagccctc acccatatga acaggaggtt tctgcgttag gtggaatacc atattctcag 4140 atatatggat ggtatcgtgt taattttggt gtgattgatg aacgattaca tcgtaacagg 4200 gaatatagag accggtatta cagaaatctg aatatagctc cggcagagga tggttacaga 4260 ttagcaggtt tcccaccgga tcaccaagct tggagagaag aaccctggat tcatcatgca 4320 ccacaaggtt gtggaaattc atcaagaaca attacaggtg atacttgtaa tgaggagacc 4380 cagaatctga gcacaatata tctcaggaaa tatcaatcaa aagttaagag gcagatattt 4440 tcagactatc agtcagaggt tgacatatat aacagaattc gggatgaatt atgaggatct 4500 gggccctaac aaaacaaaaa gatggggtta ttccctaaac ttcatgggtt acgtaattgg 4560 aagttggggg acattgccac aagatcatat tgtacaaaag atcaaacact gttttagaaa 4620 acttcctgta aacaggccta ttgattggaa agtatgtcaa aggattgtgg gtcttttggg 4680 ctttgctgct ccatttacac aatgtggata tcctgcctta atgcctttgt atgcatgtat 4740 acaagctaaa caggctttca ctttctcgcc aacttacaag gcctttctaa gtaaacagta 4800 catgaacctt taccccgttg ctcggcaacg gcctggtctg tgccaagtgt ttgctgacgc 4860 aacccccact ggctggggct tggccatagg ccatcagcgc atgcgtggaa cctttgtggc 4920 tcctctgccg atccatactg cggaactcct agccgcttgt tttgctcgca gccggtctgg 4980 agcaaagctc ataggaactg acaattctgt cgtcctctcg cggaaatata catcgtttcg 5040
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(21)P10607119-8 A2 (22) Data de Dep
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2 Cada uma destas técnicas requer
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4 construção de ácido nucleico a
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6 seqüência de gene inicial imedi
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8 codificadora operavelmente ligada
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10 expressão do HBsAg em células
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12 dominante em camundongos Balb/c.
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14 específicos de aCD4 e/ou aCD8,
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16 A SEQ ID NO: 39 é o oligonucleo
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18 Manual (2 a Edição, 1989); DNA
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20 um ácido nucleico hóspede (por
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22 imunogênico de um antígeno da
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24 regula a transcrição de um pol
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26 Nucleic Acids Research 12, p387-
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28 alguns casos as mutações podem
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30 podem ser administradas de prefe
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compreender: nativa; 32 A seqüênc
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34 (cepa Ad169 do hCMV). Uma seqü
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36 um fragmento ou variante de um p
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38 amostra do vírus, usando técni
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40 A nativo do gene inicial imediat
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5 42 Preferivelmente, o intron (c)
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44 Uma seqüência de intron heter
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46 função antiviral). A construç
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48 estimular o sistema imune do hos
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50 retroviridae (incluindo por exem
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52 80% de homologia de seqüência
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54 polipeptídeos e em particular a
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56 As construções de ácido nucle
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58 cepas da influenza diferentes. E
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60 precedentes. Em um exemplo prefe
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62 ativos das subunidades LT A e/ou
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64 Uma construção da invenção p
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66 Preferivelmente existe um aument
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68 Uma variante (ii) ou fragmento (
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70 Em um exemplo, a seqüência lí
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72 homologia de seqüência de nucl
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não traduzida. 74 Tipicamente a se
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76 Uma seqüência promotora pode s
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78 polipeptídeos. Tipicamente, a s
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mencionadas. 80 Em casos onde a con
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82 codifica um antígeno HA, uma va
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84 sejam operavelmente ligadas ao p
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86 codificadoras escolhidas em pPJV
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88 nucleico da invenção ou uma po
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90 Lipofectin , e DOTAP (1,2-bis(ol
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92 preferivelmente de 0,5 a 10 pg e
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94 partículas carreadoras (por exe
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5 96 gm, preferivelmente de cerca d
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98 podem ser selecionados para se o
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100 efeito que o elemento tem sobre
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102 menos tão altos quanto ou mais
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Materiais de partida 104 (i) pWRG71
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106 fragmento vedador que possibili
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108 PCR sem plasmídeo pFL-M2 (Joel
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110 tornar as extremidades abruptas
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Aprotinina 112 O oligo JF354 sinté
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114 pPJV7510 foi cortado com Sal 1
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116 propagadas em meio recomendado
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118 as células SCC15 são mostrado
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120 como descrito no Exemplo 9. Os
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122 Análise de Soros quanto a Anti
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124 Louis, MO)/150 mIVI de NaC1 (Fi
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Tabela 2 126 Comparação de Elemen
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128 fornecido pelo vetor de vacina
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130 que a vacina de DNA pPJV1671 in
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132 hemaglutinação (HI) na pré v
Page 135 and 136: 134 GMT da referência para cada gr
Page 137 and 138: 136 A imunização resultou em aume
Page 139 and 140: 138 indivíduos dosados com a dor b
Page 141 and 142: 140 tradução, o elemento ATG forn
Page 143 and 144: 142 antígenos virais quando co-lib
Page 145 and 146: 144 cassete de expressão B de LT f
Page 147 and 148: Números de base da seqüência em
Page 149 and 150: 148 pPJV7785 foi cortado com Xho 1
Page 151 and 152: 150 Tabela 9: Componentes de constr
Page 153 and 154: 152 et al, J. Virol. 76, 4536-4546,
Page 155 and 156: 154 populações de pérola com int
Page 157 and 158: 156 616, 1993). A comparação de s
Page 159 and 160: 158 respectivamente, ao passo que 1
Page 161 and 162: 160 codifica as subunidades A e B d
Page 163 and 164: 162 controle positive (soro contend
Page 165 and 166: 1 LISTAGEM DAS SEQÜÊNCIAS POWDER
Page 167 and 168: 3 atatatactc tatgttatac tctatgatat
Page 169 and 170: 5 30 35 40 cac cgt cc ggatctcagc aa
Page 171 and 172: 26 43 DNA Seqüência artificial
Page 173 and 174: 9 41 ggaagctagc ggcgagtgtc ccgag 2
Page 175 and 176: 11 Fusão 1/2 de exon CMV (1588).
Page 177 and 178: 13 gga agc att ccc aat gac aaa cca
Page 179 and 180: 325 330 335 Gly Met Arg Asn Val Pro
Page 181 and 182: 17 ggcgtaatgc tctgccagtg ttacaaccaa
Page 183 and 184: 19 tgt atg gaa agt gta aga aac gga
Page 185: 21 DNA Construção artificial
Page 189 and 190: Ala Arg Arg Gly Gly Glu Gln Leu Asp
Page 191 and 192: 1 REIVINDICAÇÕES 1. Construção
Page 193 and 194: 3 reivindicações de 6 a 10, carac
Page 195 and 196: imediato maior de hCMV; e 5 (a) uma
Page 197 and 198: 7 seqüência promotora e uma seqü
Page 199 and 200: imediato maior de hCMV; e 9 (b) exo
Page 201 and 202: 11 da toxina do cólera respectivam
Page 203 and 204: 13 (iii) um fragmento funcional de
Page 205 and 206: ealçadora 3'. 15 68. Construção
Page 207 and 208: 17 reivindicação 76, caracterizad
Page 209 and 210: 19 operavelmente ligado à seqüên
Page 211 and 212: fragmento; e 21 (ii) pelo menos uma
Page 213 and 214: acordo com qualquer uma das reivind
Page 215 and 216: a expressão de HBsAg ron e/ou HBV3
Page 217 and 218: 3/51 Efeito de RIA, ENH sobre a exp
Page 219 and 220: MA Enh 5/51 - Efeito de RIA, ENH so
Page 221 and 222: w 5 i g -a o o o . 'Mu/mi ou títul
Page 223 and 224: pUC19 Origem Rep 1 sítio EcoR1 RBG
Page 225 and 226: Glob pA pUC19 pUC OR HB Venh 11/51
Page 227 and 228: opu2loo042-elnooren2p4M312 plu2oReo
Page 229 and 230: Eco RJ (468L região BGHpA BOI (43
Page 231 and 232: Glob pA sgm(43 ,37) 1-18V, Nef Seq
Page 233 and 234: GIob pA pUC19 HBVenh B^HI R) RBGpA
Page 235 and 236: sítio poliA região rB-glob Pa 21/
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23/51 —0— Vermelhidão _ Incha
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VN1194H5 25/51 25/51 Figura 20 KanR
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27/51 KanR (Tn903) 601 AAATGCTTGA T
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Eagl 29/51 Exon 1/2 de CMV 1651 CCA
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BglII 31/51 LLV L W G I HHP NDA AEQ
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33/51 G V K L ESI G I Y Q I L S I Y
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35/51 4601 AAAGGCCGCG TTGCTGGCGT TT
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Sítio poliAl rGlob pA Desconhecido
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1 GGCGTAATGC TCTGCCASTG TTACAACCAA
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2101 ATGGAGTTCC GCGTTACATA ACTTACGG
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4401 TGGGGACATC ATGAAGCCCC TTGAGCAT
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pUC1 9 SapI (5247) EcaRI (5183 sít
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47/51 MATDIDMLID LGLDLSDSEL EEDALER
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C > — > 8 Ingênuo ICP27+1-7 (9:1
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a 80 C a) > .> 60 -a) Is o o a) ,se
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fe,,ijo OafffV,1- 1 PI cio 7r -- .1
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imediato maior de hCMV; e 2 (a) uma
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4 80% homologia de seqüência de a
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6 12. Construção de ácido nuclei
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das reivindicações precedentes, c
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esta; 10 (c) a seqüência do vetor
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12 (a) uma seqüência líder não
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caracterizada pelo fato de que 14 2
Page 283 and 284:
16 uma das reivindicações de 1 a
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