生物技术进展
生物技術進展
생물기술진전
2013年
2期
132-136
,共5页
苏云金芽胞杆菌%cry1Ab基因%克隆与表达%可溶性蛋白
囌雲金芽胞桿菌%cry1Ab基因%剋隆與錶達%可溶性蛋白
소운금아포간균%cry1Ab기인%극륭여표체%가용성단백
Bacillus thuringiensis%cry1Ab gene%clone and expression%soluble protein
苏云金芽胞杆菌(Bacillus thuringiensis)cry1Ab基因已广泛用于抗虫转基因植物,但其编码的杀虫蛋白对刺吸式口器害虫基本无效.本研究依据已发表cry1Ab基因序列设计一对全长引物,从Bt WB7菌株总DNA中克隆出cry1Ab基因全序列,构建原核表达载体pGEX-KG-cry1Ab并转化到大肠杆菌BL21(DE3)中,经IPTG诱导成功表达出156 kDa的Cry1Ab-GST融合蛋白.研究结果为进一步定向改造Cry1Ab使其能够正确识别刺吸式口器害虫肠道内的受体蛋白奠定基础.
囌雲金芽胞桿菌(Bacillus thuringiensis)cry1Ab基因已廣汎用于抗蟲轉基因植物,但其編碼的殺蟲蛋白對刺吸式口器害蟲基本無效.本研究依據已髮錶cry1Ab基因序列設計一對全長引物,從Bt WB7菌株總DNA中剋隆齣cry1Ab基因全序列,構建原覈錶達載體pGEX-KG-cry1Ab併轉化到大腸桿菌BL21(DE3)中,經IPTG誘導成功錶達齣156 kDa的Cry1Ab-GST融閤蛋白.研究結果為進一步定嚮改造Cry1Ab使其能夠正確識彆刺吸式口器害蟲腸道內的受體蛋白奠定基礎.
소운금아포간균(Bacillus thuringiensis)cry1Ab기인이엄범용우항충전기인식물,단기편마적살충단백대자흡식구기해충기본무효.본연구의거이발표cry1Ab기인서렬설계일대전장인물,종Bt WB7균주총DNA중극륭출cry1Ab기인전서렬,구건원핵표체재체pGEX-KG-cry1Ab병전화도대장간균BL21(DE3)중,경IPTG유도성공표체출156 kDa적Cry1Ab-GST융합단백.연구결과위진일보정향개조Cry1Ab사기능구정학식별자흡식구기해충장도내적수체단백전정기출.
The cry1Ab gene from Bacillus thuringiensis has been widely used in transgenic plants around the world. However, the Cry1Ab insecticidal protein shows no or little toxicity to the pests with piercing-suckling moutnparts. In this study, the cry1Ab gene was amplified from total DNA of Bt strains WB7 with a pair of primers designed on the full-length sequences of published cry1Ab genes. Then it was ligated with linearized pGEX-KG vector to construct recombinant expression vector pGEX-KG-cry1Ab. The soluble Cry1Ab-GST fusion protein was obtained after transferring pGEX-KG-cry1Ab into E. coli BL21 ( DE3) and then inducing by IPTG. The results provided a basis for further studies of directional modification of Cry1Ab in order that it can bind to the proper receptors in midgut of the pests with piercing-suckling moutnparts.
