苏云金芽胞杆菌cry2Ab34基因的克

发布时间：2018-01-28 19:55

本文关键词： 苏云金芽胞杆菌 cryAb基因杀虫晶体蛋白杀虫活性小菜蛾棉铃虫　出处：《生物技术通报》2017年04期 　论文类型：期刊论文

【摘要】：旨在对苏云金芽胞杆菌cry2Ab34基因进行分子鉴定和生物活测定,从杀虫活性高的Bt BJH500菌株中提取基因组DNA,通过PCR法克隆得到cry2Ab34基因(Gen Bank登录号为KX357382)。这个基因序列已被国际Bt基因命名委员会正式命名为cry2Ab34,结果显示,对cry2Ab34基因进行表达,获得大小约70 k D的表达产物,将该基因在大肠杆菌E.coli BL21中表达之后进行生物活性测定,结果表明对鳞翅目害虫小菜蛾(Plutella xyllostella)和棉铃虫(Helicoverpa armigera)都具有一定的杀虫活性,其校正死亡率10μg/m L为31.25%,100μg/m L为62.5%和校正死亡率10μg/m L为29.4%,100μg/m L为52.9%。
[Abstract]:In order to identify the cry2Ab34 gene of Bacillus thuringiensis by molecular identification and bioassay, genomic DNA was extracted from BT BJH500 strain with high insecticidal activity. Cry2Ab34 gene Gen Bank was cloned by PCR method and its accession number was KX357382. This gene sequence has been officially named cry2Ab34 by the International BT Gene Nomenclature Committee. The results showed that the cry2Ab34 gene was expressed and the expression product of 70 KD was obtained. After the gene was expressed in E. coli BL21, the bioactivity was determined. The results showed that it was effective against Plutella xyllostellaand Helicoverpa armigera (Lepidoptera) and cotton bollworm (Helicoverpa armigera). All of them have certain insecticidal activity. The corrected mortality rate of 10 渭 g / mL was 31.25 渭 g / mL, and the corrected mortality rate of 10 渭 g / mL was 62.5%, and the corrected mortality rate of 10 渭 g / mL was 29.4%. The value of 100 渭 g / mL is 52.9.
【作者单位】：东北农业大学生命科学学院;
【基金】：转基因生物新品种培育科技重大专项(2014ZX0800913B-002) 国家基础科学人才培养基金资助项目(J1210069) 国家自然科学基金项目(31401812) 黑龙江省教育厅科学技术研究项目(12521021)
【分类号】：S476.1
【正文快照】： 苏云金芽胞杆菌(Bacillus thuringiensis,Bt)是一种能够产生多种杀虫晶体蛋白(Cry)的土壤细菌[1,2],其形成芽胞的过程中,形成一个或多个伴胞晶体(parasporal crystal)。大多数这种伴胞晶体具有杀虫活性,所以又称杀虫晶体蛋白(ICPs)或δ-内毒素(δ-endotoxin)。该菌具有高效、

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