马传染性贫血病毒受体插入型基因的原核表达  

Procaryotic Expression of Inserted Receptor Gene in Equine infectious anemia virus

作　　者：杨旭艳[1,2] 张淑琴[1,2] 王雪峰[1,2] 曹贵方[1] 周建华[2] 

机构地区：[1]内蒙古农业大学动物科学与医学学院,内蒙古呼和浩特010018 [2]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

出　　处：《动物医学进展》2009年第3期36-39,共4页Progress In Veterinary Medicine

基　　金：黑龙江省发展高新技术产业专项资金项目(FW05B007);哈尔滨市科技攻关计划项目(2006AA3AS040);中国农业科学院一级岗位人才基金项目

摘　　要：采用PCR方法扩增出马传染性贫血病毒受体(EIAVR1)插入型(INR)的基因,将INR基因分别亚克隆到原核表达载体pGEX-6p-1和pET-30a中,经PCR和双酶切鉴定以及序列测定,构建其并将其转化到大肠埃希菌BL21(DE3)中进行表达。结果显示,在pET-30a-INR的表达量高于pGEX-INR,且经过优化表达条件,只在pET-30a-INR以部分可溶蛋白的形式表达,而pGEX-INR仍以包涵体形式表达;经Western blot检测,不同载体表达的蛋白均可被相应的小鼠单克隆抗体特异性识别,具有良好的抗原性。The inserted receptor （INR）gene of equine infectious anemia virus was obtained by PCR. For further study on INR, the aimed gene was cloned on the vector of pGEX-6p-1 and pET -30a. The recombinant plasmid was identified by PCR, digestion with BamHⅠ and Not Ⅰ and sequence analyse. The recombinats was transformed into E. coli BL21. The results showed that the expression level in pET- 30a - INR was higher than that in the pGEX-INR. After optimizong the condition of expression, the soluble protein was obtained in pET- 30a -INR partly. But in the pGEX-INR, only the cytorrhyctes was seen. By Western blot, the two reeombinat proteins were recognized by specific antibodies. Therefore,the antigenieity was perfect.

关 键 词：马传染性贫血病毒 受体插入型基因 原核表达 

分 类 号：S852.659.3[农业科学—基础兽医学] Q786[农业科学—兽医学]