山羊痘病毒非必需基因区gp024基因的鉴定  

Identification of non-essential gp024 gene in Goatpox virus

作　　者：雷震[1,2] 王芳[3] 马文戈[1] 黄炯[1] 符子华[1] 于力[3] 冉多良[2] 

机构地区：[1]新疆畜牧科学院兽医研究所,新疆乌鲁木齐830000 [2]新疆农业大学,新疆乌鲁木齐830000 [3]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2009年第11期842-845,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(30760181)

摘　　要：为筛选山羊痘病毒(GTPV)的外源基因插入区,本研究以GTPV Pellor株为模板,设计2对引物,PCR扩增AV41株GTPV_gp024基因相邻上下游长度约1.1kb的同源重组片段,分别插入质粒pGPT-EGFP中。插入部位在痘病毒双向启动子p11～p7.5启动的报告基因EGFP-GPT上下游,构建转移载体pEG024,并转染已感染GTPV AV41病毒的LT细胞,经GPT加压筛选7代后得到重组病毒。结果显示,重组病毒稳定表达报告基因EGFP,从而表明GTPV_gp024基因能够作为外源基因的插入区。In this study, two circumferential fragments covering 1.1 kb approximately of the locus of ＂GTPV_gp024＂ were amplified by PCR with specific primers based on GTPV strain Pellor. The PCR fragments were cloned into upstream and downstream of the enhanced green fluorescent protein （EGFP） of pGPT-EGFP vector modulated by bi-directional promoter p11-p7.5 originated from fowlpox virus. The recombinant transfer vector was co-transinfected into GTPV strain AV 41 infected LT cells. A recombinant GTPV strain was obtained after 7 rounds screening under GPT pressure. The results showed that EGFP was expressed stably by the recombinant GTPV, and the GTPV_gp024 locus could be used as a novel region for expression of foreign genes.

关 键 词：山羊痘病毒 GTPV_gp024基因座 重组病毒 

分 类 号：S852.61[农业科学—基础兽医学]