本文選題：蘇云金芽孢桿菌 + Crya基因��； 參考：《中國(guó)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(dá)系統(tǒng),通過(guò)同源重組克隆進(jìn)PHKT2表達(dá)載體,將此表達(dá)載體導(dǎo)入吡咯伯克霍爾德氏菌JK-SH007,通過(guò)誘導(dǎo)表達(dá),成功指導(dǎo)合成T7 RNA聚合酶且高效表達(dá)分子量為70k Da的Cry3a蛋白。通過(guò)粗提液對(duì)二齡榆藍(lán)葉甲進(jìn)行生物毒殺試驗(yàn),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(dá)系統(tǒng),可高效表達(dá)毒性的Cry3Aa蛋白,為進(jìn)一步開(kāi)發(fā)殺蟲生防菌劑,建立外源基因表達(dá)技術(shù)平臺(tái)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(xué)林學(xué)院;湖州師范學(xué)院;
【基金】：國(guó)家社科基金(16@ZH005) 國(guó)家自然科學(xué)基金(31550004)資助項(xiàng)目
【分類號(hào)】：Q78;S476

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 黃敏;李達(dá)旭;向文良;郭建華;趙健;張杰;楊志榮;孫群;;類產(chǎn)堿假單胞菌核心殺蟲蛋白基因的轉(zhuǎn)化及殺蟲活性[J];微生物學(xué)報(bào);2008年09期

2 潘映紅,張杰,黃大，

本文編號(hào)：1825866