本文選題：蘇云金芽孢桿菌 + Crya基因��； 參考：《中國生物工程雜志》2017年06期

【摘要】：人工合成優(yōu)化Cry3a殺蟲蛋白基因,并從大腸桿菌BL21(DE3)中克隆T7表達系統(tǒng),通過同源重組克隆進PHKT2表達載體,將此表達載體導入吡咯伯克霍爾德氏菌JK-SH007,通過誘導表達,成功指導合成T7 RNA聚合酶且高效表達分子量為70k Da的Cry3a蛋白。通過粗提液對二齡榆藍葉甲進行生物毒殺試驗,結(jié)果表明粗提液具有高毒性,致死中濃度(LC_(50)=0.63(0.48-0.82)g/L)。成功構(gòu)建的T7表達系統(tǒng),可高效表達毒性的Cry3Aa蛋白,為進一步開發(fā)殺蟲生防菌劑,建立外源基因表達技術(shù)平臺奠定了基礎(chǔ)。
[Abstract]:Cry3a insecticidal protein gene was synthesized and optimized. T7 expression system was cloned from Escherichia coli BL21 (DE3) and cloned into PHKT2 expression vector by homologous recombination. The expression vector was introduced into JK-SH007 by induction expression. T7 RNA polymerase was successfully synthesized and highly expressed Cry3a protein with a molecular weight of 70 kDa. The biological toxicity test of second instar Ulmus pumila was carried out by crude extract. The results showed that the crude extract was highly toxic, and the median lethal concentration was 0.63 ~ 0.63 ~ 0.48 ~ 0.82g / L ~ (-1) 路L ~ (-1) 路L ~ (-1) ~ (-1) ~ (-1) ~ (-1). The successfully constructed T7 expression system can express toxic Cry3Aa protein efficiently, which lays a foundation for the further development of insecticidal biocontrol agents and the establishment of foreign gene expression technology platform.
【作者單位】： 南京林業(yè)大學林學院;湖州師范學院;
【基金】：國家社科基金(16@ZH005) 國家自然科學基金(31550004)資助項目
【分類號】：Q78;S476

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