Curr. Biotech. ›› 2013, Vol. 3 ›› Issue (2): 132-136.DOI: 10.3969/j.issn.2095-2341.2013.02.10

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Cloning and Expression of cry1Ab Gene from Bacillus thuringiensis for Further Molecular Modification

CHEN Fan-bing, SHAO En-si, HUANG Zhi-peng   

  1. 1.Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education,Fujian Agriculture and Forestry University, Fuzhou 350002, China;
    2.College of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou 350002 China;
    3.College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China
  • Received:2013-03-20 Online:2013-03-25 Published:2013-03-15

用于分子改造的苏云金芽胞杆菌cry1Ab基因的克隆与表达

陈凡冰1,3,邵恩斯1,2,黄志鹏1*   

  1. 1.福建农林大学教育部生物农药与化学生物学重点实验室, 福州 350002;
    2.福建农林大学植物保护学院, 福州 350002;
    3.福建农林大学生命科学学院, 福州 350002
  • 通讯作者: 研究员,博士生导师,主要从事微生物农药研究,E-mail:zhipengh6841@yahoo.com.cn
  • 作者简介:陈凡冰,硕士研究生,研究方向为微生物农业。
  • 基金资助:

    国家自然科学基金项目(31071745);福建省自然科学基金项目(2011J01076)资助。

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Abstract: 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.

Key words: Bacillus thuringiensis, cry1Ab gene, clone and expression, soluble protein

摘要： 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。

关键词: 苏云金芽胞杆菌, cry1Ab基因, 克隆与表达, 可溶性蛋白

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