禽流感病毒AM2蛋白相互作用宿主蛋白的筛选及初步验证
发布时间：2019-01-01 18:33
【摘要】：采用酵母双杂交系统筛选与禽流感病毒AM2相互作用的宿主蛋白,可为研究AM2蛋白的生物学新功能提供依据。将检测无毒性与自激活作用的诱饵质粒pGBKT7-AM2与淋巴文库质粒进行融合杂交,然后对经选择性培养基筛选到的阳性克隆进行酶切鉴定和测序分析,阳性克隆进行酵母回转试验验证。经酵母双杂交筛选与验证最终获得2个与AM2相互作用的蛋白。选择线粒体细胞色素C氧化酶亚单位Ⅱ(COXⅡ)蛋白进行进一步研究,构建真核表达载体pcDNA3.0-HA-COXⅡ,并进行GST-pull down和免疫荧光试验验证。结果表明,COXⅡ与AM2存在相互作用,且在细胞质中存在共定位现象,为进一步研究禽流感病毒的致病机理和病毒-宿主相互作用机制提供了理论基础。
[Abstract]:Yeast two-hybrid system was used to screen host proteins interacting with AM2 of avian influenza virus, which could provide basis for the study of new biological functions of AM2 protein. The bait plasmid pGBKT7-AM2, which is nontoxic and self-activated, was fused with the lymphoid library plasmid, and the positive clones screened by selective culture medium were digested and sequenced. The positive clones were tested by yeast rotation test. Two proteins interacting with AM2 were obtained by yeast two-hybrid screening and verification. The mitochondrial cytochrome C oxidase subunit 鈪，
本文编号：2397951
[Abstract]:Yeast two-hybrid system was used to screen host proteins interacting with AM2 of avian influenza virus, which could provide basis for the study of new biological functions of AM2 protein. The bait plasmid pGBKT7-AM2, which is nontoxic and self-activated, was fused with the lymphoid library plasmid, and the positive clones screened by selective culture medium were digested and sequenced. The positive clones were tested by yeast rotation test. Two proteins interacting with AM2 were obtained by yeast two-hybrid screening and verification. The mitochondrial cytochrome C oxidase subunit 鈪，
本文编号：2397951
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