鸭瘟病毒强、弱毒株TK基因的克隆与生物信息学分析
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摘要
本研究根据Gen Bank上发表的鸭瘟病毒(DPV)基因序列设计了1对引物,分别对DPV AV 1221株和DPV鸡胚化弱毒疫苗株的TK基因进行扩增,扩增的目的片段克隆至载体p MD19-T,经PCR和双酶切(Bam HⅠ+HindⅢ)鉴定,将阳性质粒进行测序和生物信息学分析。结果成功克隆出了长度均为1 077 bp的DPV TK基因,生物信息学分析结果表明两个片段的同源性达到100%。
According to the sequence of TK gene in Gen Bank,a pair of primers was designed to amplify this gene in virulent and vaccine duck plague virus strains. The purpose amplified fragment was cloned into p MD 19-T vector. The recombinant expression vector was selected and identified by PCR and restriction enzyme digestion(Bam HⅠ+HindⅢ). The positive recombinant plasmid was then sequenced and the biological information was also analyzed. The results showed this research successfully cloned the DPV TK gene,and both of the fragments were 1 077 bp,the result of bioinformatics analysis revealed the homology of two fragments up to 100%.
According to the sequence of TK gene in Gen Bank,a pair of primers was designed to amplify this gene in virulent and vaccine duck plague virus strains. The purpose amplified fragment was cloned into p MD 19-T vector. The recombinant expression vector was selected and identified by PCR and restriction enzyme digestion(Bam HⅠ+HindⅢ). The positive recombinant plasmid was then sequenced and the biological information was also analyzed. The results showed this research successfully cloned the DPV TK gene,and both of the fragments were 1 077 bp,the result of bioinformatics analysis revealed the homology of two fragments up to 100%.
引文
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[3]葛菡,程安春,汪铭书,等.疱疹病毒TK基因的研究进展[J].中国兽医科学,2008,38(1):86-90.
[4]樊振华,孟帆,吴忻,等.猪伪狂犬病病毒山西株胸苷激酶基因序列分析[J].中国畜牧兽医,2014,41(4):12-17.
[5]Kit S.Thymidine kinase[J].Microbiological Science,1985,12(2):369-375.
[6]李昌主,韩先杰,邹玲,等.鸭瘟病毒AV 1221 TK基因的克隆与序列分析[J].青岛农业大学学报,2007,24(3):162-164.
[7]宋得华,潘华奇,黎应胜,等.鸭瘟病毒TK基因及其编码蛋白的生物信息学分析[J].安徽农业科学,2007,35(31):9 935-9 936.
[8]葛菡,徐超,程安春,等.鸭瘟病毒TK基因的克隆及其分子特性分析[J].中国兽医科学,2008,38(4):297-302.
[2]Gale M J,Tan S L,Katze M G.Translational control of viral gene expression in eukaryotes[J].Microbiology and Molecular Biology Review,2000,64(2):239-280.
[3]葛菡,程安春,汪铭书,等.疱疹病毒TK基因的研究进展[J].中国兽医科学,2008,38(1):86-90.
[4]樊振华,孟帆,吴忻,等.猪伪狂犬病病毒山西株胸苷激酶基因序列分析[J].中国畜牧兽医,2014,41(4):12-17.
[5]Kit S.Thymidine kinase[J].Microbiological Science,1985,12(2):369-375.
[6]李昌主,韩先杰,邹玲,等.鸭瘟病毒AV 1221 TK基因的克隆与序列分析[J].青岛农业大学学报,2007,24(3):162-164.
[7]宋得华,潘华奇,黎应胜,等.鸭瘟病毒TK基因及其编码蛋白的生物信息学分析[J].安徽农业科学,2007,35(31):9 935-9 936.
[8]葛菡,徐超,程安春,等.鸭瘟病毒TK基因的克隆及其分子特性分析[J].中国兽医科学,2008,38(4):297-302.