新城疫病毒鹌鹑分离株融合蛋白基因的克隆与序列分析  

Sequencing of Fusion Glycoprotein Gene of Newcastle Disease Virus Isolated from Quail

作　　者：郭鑫[1] 曹殿军[1] 闵平[1] 闫丽辉[1] 刘培欣[1] 房海[2] 卢景良[1] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所,哈尔滨150001 [2]河北农业技术师范学院

出　　处：《中国预防兽医学报》1999年第6期430-435,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家攀登计划B项目!85-44-01-44;自然科学基金!39893290-4

摘　　要：本研究以一株从自然发生新城疫病死的鹌鹑中所分离到的新城疫病毒（QNDV／89）为研究对象，用反转录一聚合酶链式反应（RT-PCR）对QNDV／89株的F全基因进行分段扩增、定向克隆到pUC119载体后，采用Sanger’s双脱氧末端终止法测定其核苷酸序列，并推导出相应氨基酸序列。QNDV／89株F基因全长1662bp，单一的开放阅读框编码553个氨基酸的多肽，其裂解位点的氨基酸序列为112R-R-Q-R-R-F117，是NDV强毒株所特有的序列结构，这与其生物学特性及致病性实验结果是相一致的。序列分析结果表明，QNDV／89株与F48E9株的核苷酸同源率高达99.22％，氨基酸的同源率为98.37％;而与弱毒株LaSota的核苷酸同源率为88.93％，氨基酸的同源率为90.42％。本研究首次报道了流行于我国鹌鹑中的新城疫强毒融合蛋白基因序列，并与其它新城疫病毒进行了同源性比较，证明QNDV／89株核苷酸的变异引起了其蛋白质二级结构的部分改变。A Newcastle disease virus(NDV ) , QNDV/89, isolated from a quail died of disease was investigated. According to published NDV fusion protein (F) gene sequence, two pairs of specific primers were designed and synthesized. Full-length F gene was amplified by reverse transcription and polymerase chain reaction (RT-PCR) and cloned into pUC119. The gene was sequenced by Sanger's method and open reading frame of 1662 bp was determined, and the cleavage site on the deduced deptide was112R-R-Q-R-R-F117, indicated it's a typical virulent NDV strain. The sequence comparison showed 99. 22% homology in nucleotide and 98. 37% in amino acid with F48E9,while compared with La Sota thers are 88. 93% and 90. 42% respectively. This study demonstrated that the nucleotide variations of QNDV/89 strain may cause the change of secondary structure of F protein.

关 键 词：新城疫病毒 鹌鹑 分离株 融合蛋白基因 克隆 

分 类 号：S852.65[农业科学—基础兽医学]