cry1Ac基因啟動子表達Vip3Aa蛋白特性分析
發(fā)布時間:2018-05-22 11:38
本文選題:蘇云金芽胞桿菌 + cryAc啟動子; 參考:《東北農(nóng)業(yè)大學學報》2017年09期
【摘要】:蘇云金芽胞桿菌cry基因啟動子常用于構(gòu)建蛋白表達載體。為探討蘇云金芽胞桿菌cry基因啟動子指導Vip3Aa蛋白表達情況及殺蟲活性,以p UC19載體為基礎(chǔ),運用重疊引物PCR方法構(gòu)建Vip3Aa11表達載體,并與由T7啟動子指導的Vip3Aa11表達蛋白殺蟲活性、抗胰蛋白酶穩(wěn)定性比較,初步探索發(fā)酵條件。結(jié)果表明,cry1Ac啟動子與T7啟動子均在上清液中表達大小為88 ku Vip3Aa11蛋白,對甜菜夜蛾、棉鈴蟲殺蟲活性差異不顯著,cry1Ac基因啟動子在37℃、48 h更適合Vip3Aa11蛋白的表達,為vip基因表達、功能驗證及殺蟲機理等研究提供新思路。
[Abstract]:The promoter of cry gene of Bacillus thuringiensis is often used to construct protein expression vector. In order to investigate the expression and insecticidal activity of cry gene promoter of Bacillus thuringiensis, based on p UC19 vector, the Vip3Aa11 expression vector was constructed by using overlapping primer PCR method, and the insecticidal activity of Vip3Aa11 expression protein guided by T7 promoter was obtained. The stability of antitrypsin was compared and the fermentation conditions were preliminarily explored. The results showed that both cry1Ac promoter and T7 promoter expressed 88 ku Vip3Aa11 protein in supernatant. The difference of insecticidal activity of Helicoverpa armigera was not significant. Cry1Ac promoter was more suitable for the expression of Vip3Aa11 protein at 37 鈩,
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