禽流感病毒AM2蛋白相互作用宿主蛋白的篩選及初步驗證
發(fā)布時間：2019-01-01 18:33
【摘要】：采用酵母雙雜交系統(tǒng)篩選與禽流感病毒AM2相互作用的宿主蛋白,可為研究AM2蛋白的生物學新功能提供依據(jù)。將檢測無毒性與自激活作用的誘餌質(zhì)粒pGBKT7-AM2與淋巴文庫質(zhì)粒進行融合雜交,然后對經(jīng)選擇性培養(yǎng)基篩選到的陽性克隆進行酶切鑒定和測序分析,陽性克隆進行酵母回轉(zhuǎn)試驗驗證。經(jīng)酵母雙雜交篩選與驗證最終獲得2個與AM2相互作用的蛋白。選擇線粒體細胞色素C氧化酶亞單位Ⅱ(COXⅡ)蛋白進行進一步研究,構建真核表達載體pcDNA3.0-HA-COXⅡ,并進行GST-pull down和免疫熒光試驗驗證。結(jié)果表明,COXⅡ與AM2存在相互作用,且在細胞質(zhì)中存在共定位現(xiàn)象,為進一步研究禽流感病毒的致病機理和病毒-宿主相互作用機制提供了理論基礎。
[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，
本文編號：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，
本文編號：2397951
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