PCR快速克隆猴免疫缺陷病毒核心蛋白基因  

RAPID CLONING OF SIV CORE PROTEIN GENE WITH POLYMERASE China REACTION

作　　者：何伏秋[1] 施慧君[1] 潘勇[1] 丛喆[1] 吴小闲[1] 

机构地区：[1]中国协和医科大学实验动物研究所,中国医学科学院北京100021

出　　处：《中国人兽共患病杂志》1998年第5期30-33,共4页Chinese Journal of Zoonoses

摘　　要：SIV为猴艾滋病的病原，其核心蛋白基因保守性强，它编码的27kD蛋白是病毒颗粒的主要结构蛋白，可做为SIV感染的主要诊断依据。我们根据SIV基因序列，设计合成了一对含有两个合适酶切位点的特异性PCR引物；PCR法直接从SIVmac毒株感染的恒河猴外周血淋巴细胞基因组DNA中扩增分离SIV病毒核。心蛋白编码区段，扩增产物经纯化和EcoRI、Sall双酶切后与相同酶切位点的质粒载体pBV220连接，转化大肠杆菌DH52，经巢式PCR快速筛选，获得了SIV核心蛋白基因重组质粒pBVSG；核苷酸序列测定发现了7处碱基点突变，作者分析这是由于SIVmac毒株在中国恒河猴体内感染适应后发生基因突变所致。Simian immunodeficiency virus (Sly) causes simian immunodeficiency disease syndrome(SAIDS),its core protein gene encoding 27KD protein is very conservative,the 27KD protein is one of SIVstruc-tive proteins,and it can be used as diagnostic base of SIV infection.. Two specific primers corresponding to SIVgene sequence were designed and synthesized. PCR directly ampified the SIV core protein gene from lymphocytecell genome of SIV-infected Rhesus monkeys. The purified PCR product was inserted into vector pBV220 afterdi-gestion with EcoRI and Sall,then the inserts were transmitted into Ecoli. DH52. The specific recombinants werescreened by nested PCR. We have succeeded in obtaining SIV core protein gene recombinant plasmid named pB-VSG. pBVSG was sequenced and compared with SIVmac251,seven gene mutation sites were founded in the region ofSIV core protein gene,it maybe result from the mechanism of viral adaptation during SIV infected monkeys.

关 键 词：SIV PCR 巢式PCR 免疫缺陷病毒 猴 

分 类 号：R512.91[医药卫生—内科学] R373.9[医药卫生—临床医学]