新城疫病毒V4株主要基因的克隆与序列分析  

Cloning and sequencing of majority gene in Newcastle disease virus V4 strain

作　　者：刘芳宁[1] 闫丽辉[1] 曹殿军[1] 刘培欣[1] 杨增岐[2] 

机构地区：[1]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室,黑龙江哈尔滨150001 [2]西北农林科技大学动物科技学院,陕西杨凌712100

出　　处：《中国兽医学报》2007年第3期319-321,362,共4页Chinese Journal of Veterinary Science

基　　金：国家重点基础研究发展规划"973"项目(G1999011902);黑龙江省科学技术计划资助项目(GB02B505)

摘　　要：设计4对引物，利用RT—PCR技术分别扩增出新城疫病毒（NDV）V4株长约4.2、4.1、4.1、3.5kb的4个互相重叠的核苷酸片段，并分别进行了克隆和序列测定。将这4个片段的核苷酸序列拼接后，得到长为15 061 bp的核苷酸序列。该序列包含有NDVv4株结构基因NP的全部编码区和5’端非编码序列，结构基因P、M、F、HN及L基因的全部编码区和非编码区序列，各结构基因之间的间隔序列以及基因组5’端部分尾巴序列（108bp）。序列比较分析表明，NDVv4株F蛋白裂解位点序列为^112GKQGRL^117，HN基因编码616个氨基酸，符合无毒株的特征；NDVv4株同基因组长度为15 192 bp的毒株一样，在基因组的1647～1648bp间比基因组长为15192bp的毒株少了6个碱基。A pair of primers were designed. Utilizing RT-PCR, 4 overlapping cDNA fragments of NDV V4 strain were successfully amplified using above NDV specific primers. The 4 PCR fragments length is about 4.2, 4.1,4.1,3.5 kb respectively,which were cloned and sequenced. The nucleotide sequence of 15 061 bp in length gained by spliced the nucleotide sequence of the 4 fragments, which covers the part of nucleocappsid protein（NP）, the whole nucleotide sequence of Phosp hoprotein （P）, Matrix protein （M）, Fusion pro tein （F）, Haemaggl u tinin-Neu- raminidase（HN） ,Large protein（L）,the intergenic regions in NP-P,P-M,M-F,F-HN,HN-L junctions,and the 108 bp of 5＇ trailer region of NDV V4 strain. But the gained NP nucleotide sequence have spanned its open reading frame （ORF） and 3＇ non-coding region. According to the length of complete genome sequence known at present,we found there are at least two types of NDV strains..15 186 bp and 15 192 bp. Equaled to the 15 186 bp strain,the NDV V4 strain isn＇t the extra 6 nucleosides insertion between 1 647 nt and ] 652 nt as compared with the 15 192 bp strain. The amino acid sequence of F protein cleavage site is n2GKQGRLn7 ,and the amino acid sequence of HN protein is 616 in length,which conformed to the trait of lentogenic strain.

关 键 词：新城疫病毒 主要基因 克隆 序列分析 

分 类 号：Q78[生物学—分子生物学] S852.659[农业科学—基础兽医学]