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Position: Home > Articles > Procaryotic Expression of Inserted Receptor Gene in Equine infectious anemia virus Progress in Veterinary Medicine 2009,30 (3) 40-43

马传染性贫血病毒受体插入型基因的原核表达

作  者：
杨旭艳;张淑琴;王雪峰;曹贵方;周建华
单  位：
内蒙古农业大学动物科学与医学学院;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
关键词：
马传染性贫血病毒;受体插入型基因;原核表达
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。
译  名：
Procaryotic Expression of Inserted Receptor Gene in Equine infectious anemia virus
作  者：
YANG Xu-yan1,2,ZHANG Shu-qin1,2,WANG Xue-feng1,2,CAO Gui-fang1,ZHOU Jian-hua2(1.College of Animal Science and Veterinary Medicine,Inner Mongolia Agricultural University,Huhhot, Inner Mongolia,010018,China;2.National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin,Heilongjiang,150001,China)
关键词：
EIAVR1 inserted receptor gene;prokaryotic expression
摘  要：
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 NotⅠand 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 recombinat proteins were recognized by specific antibodies. Therefore,the antigenicity was perfect.

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