新城疫病毒贵州不同鸡源分离株F基因的克隆与序列分析

应用PCR技术扩增NDV贵州不同鸡源分离株,包括肉鸡源P1株与BY株、蛋鸡源H2株与FW株、七彩山鸡源N98株和越南斗鸡源DQ株的F蛋白基因,将该基因片段分别进行克隆和测序,并与国内外NDV参考株的对应序列进行比较分析。结果表明:6株NDV贵州不同鸡源分离株的F基因长度均为1 662 bp,编码553个氨基酸;分离株间核苷酸同源性为86.9%~99.7%,氨基酸同源性为91.0%~99.3%;与国内外NDV代表株(LaSota株、B1株、F48E9株、CH2000株和TW 2000株)的核苷酸同源性为84.0%~98.9%,氨基酸同源性为87.2%~98.6%;经系统发育进化树分析,DQ株、N98株、P1株和H2株为基因VIId型,而BY株和FW株为基因IX型。这些结果提示贵州省不同鸡群间存在相同NDV毒株感染的可能性,不同年份间NDV毒株发生基因型改变,而近年来贵州省流行的ND疫情主要是由NDV基因VII型引起。

Cloning and Sequencing of Fusion Glycoprotein Gene of Different Newcastle Disease Virus Strains Isolated from Chickens in Guizhou

The fusion protein gene(F gene) of 6 isolates of Newcastle disease virus(NDV) from different avian in Guizhou province were amplified using polymerase chain reaction(PCR) method,then cloned into the pMD18-T vector,respectively.And these F gene were sequenced and analyzed with reference NDV stains in GenBank.The results showed that the F gene of all 6 isolates are 1662 bp and encoding 553 amino acids.The sequence identity matrix of 6 isolates indicated that the homology of nucleotide sequence are 86.9%~99.7% and that of amino acid sequence are 91.0%~99.3%.The homology of nucleotide and amino acid sequence of 6 isolates,compared with that of Lasota,B1,F48E9,CH2000 and TW2000,are 84.0%~98.9% and 87.2%~98.6% respectively.The phylogenetic analysis showed that strain DQ,N98,P1 and H2 belonged to VII genotype and strain BY and FW to IX genotype.These results revealed that the same genotype NDV could induce outbreaks in different avian and the genotype of NDV had transformed in different year,and VII genotype is the main genotype of NDV occurred in Guizhou province now.