摘要
目的对广西新分离乙脑病毒GP0722株进行全基因序列测定和分析,了解其基因组结构及毒力特征。方法应用乙脑病毒全基因组扩增引物进行RT-PCR扩增,PCR产物直接测序,拼接后得到全基因序列。应用Clustal X(1.8)、DNASTAR、Mega 4.1等生物软件进行核苷酸序列及氨基酸序列分析和病毒的系统进化分析。结果广西新分离乙脑病毒GP0722全基因长10 965个核苷酸,从97到10 395位编码一个开放阅读框,编码3 432个氨基酸,与目前使用的减毒活疫苗株SA-14-14-2株比较,只有88.9%的核苷酸同源性,97.6%的氨基酸同源性,全基因组共存在1 222个核苷酸差异,83个氨基酸差异。与GenBank中选择的21株乙脑病毒全基因序列比较发现,其核苷酸总体差异率为0.9%～18.8%,氨基酸总体差异率为0.1%～5.2%。通过PrM/C区段、E区段、3′NTR区段和全基因序列进行系统进化分析显示该毒株属于基因1型乙脑病毒。结论新分离的乙脑病毒GP0722株属于基因1型,与JEV/sw/Mie/40/2004进化关系最近,与疫苗株SA-14-14-2相比关键位点氨基酸未见变异,现行使用的疫苗仍能保护GP0722引起的感染。
In order to sequence and analyze the complete nucleotide sequence of Japanese encephalitis virus（JEV） strain GP0722 newly isolated in Guangxi province,and to provide information for the genomic structure and characteristics of virulence.RT-PCR was used to amplify the sequenced fragments directly and bioinformatics softwares were used to analyze the nucleotide,amino acid and phylogenetic trees of the strain including ClustalX（1.8）,DNAStar,Mega（4.1）.The result of sequence analysis showed that the genome of GP0722 strain was 10 965 nucleotide long.An open reading frame from 97 to 10 395 was found capable of coding for a 3 432 amino acid polyprotein.Compared with the live attenuated vaccine strain SA14-14-2 in China,there were 1 222 nucleotide differences and 83 amino acid divergence.Comparison of the complete genome sequences of different JEV showed a 0.9%-18.8% nucleotide sequence divergence among them,which resulted in 0.1%-5.2% amino acid sequence divergence.Phylogenetic analysis through PrM/C,E,3′NTR and full-length genome showed that the GP0722 strain belonged to genotype 1.In conclusion,the GP0722 strain isolated in Guangxi belonged to genotype 1 and is close to the JEV/sw/Mie/40/2004.There are no mutations.Vaccination can protect human from the infection of GP0722.
出处
《中国人兽共患病学报》
CAS
CSCD
北大核心
2012年第3期270-273,共4页
Chinese Journal of Zoonoses
基金
广西壮族自治区卫生厅计划课题(No.Z2008204)
关键词
乙型脑炎病毒
基因
种系发生
Japanese encephalitis virus
genes
phylogeny
