增强型绿色荧光蛋白eGFP基因与犬瘟热N蛋白基因重组质粒的构建  

Construction of Recombinant Plasmid with Enhanced Green Fluorescent Protein Gene eGFP and N Protein Gene of Canine Distemper Virus

作　　者：夏婷婷[1] 苏晓慧[1] 翟少华[1] 简子健[1] 

机构地区：[1]新疆农业大学动物医学学院,乌鲁木齐830052

出　　处：《新疆农业科学》2015年第2期349-353,共5页Xinjiang Agricultural Sciences

基　　金：国家自然科学基金项目(31360623);新疆维吾尔自治区普通高校重点学科(基础兽医学)科研启动基金

摘　　要：【目的】构建增强型绿色荧光蛋白(e GFP)基因与犬瘟热核蛋白N基因(CDV N)的重组质粒,为获得犬瘟热重组病毒的感染性克隆奠定研究基础。【方法】设计重组PCR引物,分别扩增e GFP,CDV N基因,并融合重组基因。【结果】成功克隆了e GFP基因和CDV N基因,大小分别为720 bp和1 572 bp,经测序鉴定与Gen Bank中已发表的e GFP(登录号:X83959)和CDV N(登录号:AY466011)序列同源性分别为100%和99.81%。重组基因e GFP-CDV N大小为2 292 bp,与预期大小一致。【结论】采用重组PCR技术构建的Te GFP-CDVN重组质粒,可用于e GFP-CDVN重组蛋白的表达。【Objective】The recombinant plasmid of enhanced green fluorescent protein gene e GFP and N protein gene in canine distemper virus was constructed for the infectious clone of recombinant canine distemper virus in this paper.【Method】The recombinant PCR primers for e GFP gene and CDV N gene were designed and amplified. The amplified products were fused again by recombinant PCR.【Result】The fragments of e GFP gene and CDV N gene were 720 bp and 1 572 bp,and homology were 100% and 99. 81%,respectively,compared with the Gen Bank e GFP（ ID： X83959） and CDV N（ ID：AY466011）. The size of fusion gene e GFP-CDV N was 2 292 bp,consistently with the expected one.【Conclusion】The recombinant plasmid of T- e GFP- CDVN was successfully constructed. It can be used for the expression of fusion gene e GFP- CDV N.

关 键 词：重组PCR EGFP基因 犬瘟热N蛋白 重组基因 

分 类 号：S852.65[农业科学—基础兽医学]