| 日本乙型脑炎病毒分离株E基因的克隆及序列分析 |
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| 引用本文： | 陈梅,王艳允.日本乙型脑炎病毒分离株E基因的克隆及序列分析[J].安徽农业科学,2012(13):7746-7748,7772. |
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| 作者姓名： | 陈梅 王艳允 |
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| 作者单位： | 南京师范大学生命科学学院,江苏南京,210046 |
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| 摘 要： | 目的]对日本乙型脑炎病毒分离株E基因进行克隆及序列分析。方法]根据乙型脑炎病毒SA14和SA14-14-2株序列,设计并合成一对E基因的特异性引物,用RT-PCR方法扩增出JEV分离株E基因片段,将其与pMD19-T载体连接,应用DNAStar、Clustal_1.81和Mega序列分析软件进行同源性比较。结果]分离株E基因的cDNA长1 500 bp,可编码500个氨基酸;分离株E基因与VN50/Viet Nam/1989/Hu-man brain株、SA14株、Whe株、Benjing 1株、P3株、KV1889株J、aGAr01株和SA14-14-2株的核苷酸同源性为88.0%～99.1%,氨基酸同源性为97.8%～99.8%;分离株为乙型脑炎病毒强毒株。结论]该研究为乙脑病毒基因工程疫苗的研发奠定了一定的基础。
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| 关 键 词： | 乙型脑炎病毒 分离株 E基因 克隆 序列分析 |
Cloning and Sequence Analysis of E Genes of Japanese Encephalitis Virus Isolated Strain |
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| Institution: | CHEN Mei et al(College of Life Science,Nanjing Normal University,Nanjing,Jiangsu 210046) |
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| Abstract: | Objective] The aim was to clone and analyze the sequence of E genes of Japanese encephalitis virus isolated strain.Method] A pair of specific primers of E genes were designed according to the gene sequence of JEV SA14 and SA14-14-2 strain.The specific fragment of isolated strain E genes had been successfully amplified by RT-PCR and sequenced after being cloned directly into the pMD19-T.The homology of nucleotide and deduced amino acid between E genes with different strains which shown on GenBank by the softwares of DNAStar、Clustal1.81 and Mega was compared.Result]The results showed that the cDNA of E genes was 1 500 bp,and the homology of 88.0%-99.1% in nucleotide and 97.8%-99.8% in amino acid with the VN50/Viet Nam/1989/Human brain,SA14,Whe,Benjing 1,P3,KV1889,JaGAr01 and SA14-14-2 strain.The isolated strain was a virulent strain.Conclusion] The study lays a foundation for research and development of the genetic engineering vaccine of Japanese encephalitis virus. |
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| Keywords: | Japanese encephalitis virus Isolated strain E gene Cloning Sequence analysis |
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