新城疫新型粘膜疫苗及其构建方法

摘要

新城疫新型粘膜疫苗及其构建方法，涉及一种新城疫疫苗以及构建的方法，先通过反转录—聚合酶链式RT－PCR反应，扩增出新城疫病毒融合蛋白F基因；再将F基因与质粒pVAX1进行连接反应，构建DNA疫苗pVAX1－F；最后将质粒pVAX1－F转化进减毒鼠伤寒沙门氏菌SL7207中，最终获得DNA长度为4647bp的新城疫病毒DNA疫苗。本发明采用以减毒鼠伤寒沙门氏菌运送新城疫病毒DNA疫苗，构建新城疫新型粘膜疫苗SL7207(pVAX1－F)。通过口服方式(饮水或拌料)对家禽(鸡、鹅、鸽等)直接进行免疫，较传统的新城疫弱毒疫苗安全，且较传统的新城疫油乳剂灭活疫苗使用方便，可克服母源抗体干扰以及油乳剂灭活疫苗不能产生粘膜免疫应答的缺陷，并能提供对新城疫病毒强毒攻击的保护。

主权项

1、新城疫新型粘膜疫苗，其特征在于其DNA疫苗的DNA长度为4647bp，其DNA排序为：gactcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta 60atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata 120acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat 180aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga 240ctatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc 300ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt 360atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat 420gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag 480tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc 540aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga 600ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga 660aattaatacg actcactata gggagaccca agctggctag cgtttaaact taagcttcaa 720tatgggctcc agaccttcta ccaggatccc agcacctctg atgctgatca cccggattat 780gctgatgttg ggctgtatcc gtccgacaag ctctcttgac ggcaggcctc ttgcagctgc 840aggaattgta gtaacaggag ataaggcagt caatgtatac acctcgtctc agacagggtc 900aatcatagtc aagttgctcc cgaatatgcc cagggataag gaagcgtgtg cgaaagcccc 960attagaggca tataacagaa cactgactac tttgctcact cctcttggcg actccatccg 1020caagatccaa gggtctgtgt ccacgtctgg aggaaggaga caaaaacgct ttataggtgc 1080tattattggc agtgtagctc ttggggttgc aacagcggca cagataacag cagctgcggc 1140cctaatacaa gccaaccaga atgctgccaa catcctccgg cttaaggaga gcattgctgc 1200aaccaatgaa gctgtgcatg aagtcaccga cggattatca caactatcag tggcagttgg 1260gaagatgcag cagtttgtca atgaccagtt taataataca gcgcgagaat tggactgtat 1320aaaaatcaca caacaggttg gtgtagaact caacctatac ctaactgaat tgactacagt 1380attcgggcca cagatcacct cccctgcatt gactcagctg accatccagg cactttataa 1440tttagctggt ggcaatatgg attacttatt aactaagtta ggtataggga acaatcaact 1500cagctcatta attggtagcg gcctgatcac tggttaccct attctgtacg actcacagac 1560tcaactcttg ggcatacaag tgaatttgcc ctcagtcggg aacttaaata atatgcgtgc 1620cacctatttg gagaccttat ctgtaagtac aaccaaagga tatgcctcag cacttgtccc 1680gaaagtagtg acacaagtcg gttctgtgat agaagagctt gacacctcat actgtataga 1740gtccgatctg gatttatatt gtactagaat agtgacattc cccatgtccc caggtattta 1800ttcctgtttg agcggcaaca catcagcttg catgtattca aagactgaag gcgcactcac 1860cacgccgtat atggccctta aaggctcagt tattgccaat tgtaagataa caacatgtag 1920atgtacagac cctcctggta tcatatcgca aaattatgga gaagctgtat ccctgataga 1980tagacattcg tgcaatgtct tatcattaga tgggataact ctgaggctca gtggggaatt 2040tgatgcaact tatcaaaaga acatctcaat actagattct caagtcatcg tgacaggcaa 2100tcttgatata tcaactgaac ttggaaacgt caacaattca atcagcaatg ccttggataa 2160gttggcggaa agcaacagca agctagaaaa agtcaatgtc agactaacca gcacatctgc 2220tctcattacc tatattgttc taactgtcat ttctctagtt ttcggtgcac ttagtctggg 2280tttagcgtgt tacctgatga tcaaacagaa ggcacaacaa aagaccttgc tatggcttgg 2340gaataatacc ctcgatcaga tgagagccac tacaagagca tgaatgcaga taagaggtgg 2400gaattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgtttaaacc 2460cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2520gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2580attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2640agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2700gcttctactg ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct 2760ctggtaaggt tgggaagccc tgcaaagtaa actggatggc tttctcgccg ccaaggatct 2820gatggcgcag gggatcaagc tctgatcaag agacaggatg aggatcgttt cgcatgattg 2880aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg 2940actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg 3000ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg 3060aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg 3120ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc 3160tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc 3240tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc 3300gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc 3360aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg 3420atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct 3480tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt 3540tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc 3600tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt 3660tcttctgaat tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt 3720gcggtatttc acaccgcata caggtggcac ttttcgggga aatgtgcgcg gaacccctat 3780ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata 3840aatgcttcaa taatagcacg tgctaaaact tcatttttaa tttaaaagga tctaggtgaa 3900gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 3960gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 4020ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 4080gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 4140ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 4200cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 4260cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 4320ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 4380tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 4440cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 4500ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 4560aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctgggctt 4620ttgctggcct tttgctcaca tgttctt。