摘要
为比较NDV江苏分离株的遗传变异特征,应用RT-PCR扩增出新城疫病毒(NDV)江苏徐州株(JSXZ)、江苏宿州株(JSSZ)、江苏连云港株(JSLYG)和江苏淮安株(JSHA)的融合蛋白F基因的全长核苷酸,将其分别克隆至pMD18-T载体中。将获得的阳性重组质粒进行序列测定后,推导出其氨基酸序列,进行分析和比较。结果表明,所获得的4个分离株F基因完整的开放阅读框长度为1662bp,共编码F蛋白的553个氨基酸;4个毒株与常用疫苗株之间核苷酸序列同源性只有69.3%~80.8%。JSXZ株和JSSZ株F基因的裂解位点为112-RRQK/RRF-117,符合强毒株裂解区氨基酸组成的特征。以1bp^374bp的核苷酸序列绘制系统发育树分析表明,JSXZ和JSSZ株NDV为基因Ⅷ型,JSLYG和JSHA株NDV为基因Ⅸ型。
In order to understand the features of hereditary variation of NDV isolates from Jiangsu, the fusion glyco- protein （F） genes ofNDV Xuzhou （JSXZ） , Suzhou （JSSZ） , Lianyungang （JSLYG） and Huaian1 （JSHA） isolates were amplified by reverse transcriptase polymerase chain reaction （RT-PCR） and cloned into pMD 18-T vector. The recombinant plasmids were sequenced. Their eDNA were obtained and their amino acid sequences were deduced, analyzed and compared. The results showed that complete open read frame of F gene of the 4 isolates were 1662 bp and encoded 553 amino acids. The nucleotide sequence homologies among the 4 isolates and NDV vaccine strain were 69.3% -80.8%. The deduced amid acid sequence of the cleavage site region of JSXZ and JSSZ F protein was 112-RRQK/RRF-117, conforming to the feature of velogenic strain. Polygenetic tree of NDV strains were constructed by nucleotide acid se- quences of a 374 bp region ofF gene and showed that JSXZ and JSSZ isolates belonged to the genetype Ⅷ, JSLYG and JSHA isolates belong to the genetype Ⅸ.
出处
《中国动物检疫》
CAS
2014年第3期64-67,共4页
China Animal Health Inspection
基金
山东省优秀中青年科学家科研奖励基金(BS2011SW026)
关键词
NDV
F基因
克隆
序列分析
Newcastle disease virus
F gene
Cloning
sequence analysis
