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中国农学通报

中国农学通报 ›› 2011, Vol. 27 ›› Issue (7): 222-226.

所属专题： 生物技术； 油料作物

• 生物技术科学 • 上一篇    下一篇

苏云金芽孢杆菌cry1基因的克隆、植物表达载体的构建及转化大豆

冯頔 张莉弘 魏毅 刘金亮 潘洪玉 张世宏   

  • 收稿日期:2010-12-10 修回日期:2011-01-04 出版日期:2011-04-05 发布日期:2011-04-05
  • 基金资助:

    农业部转基因生物新品种培育重大专项

Cloning, Construction of Plant Expression Vector and Establishment of Transformation System in Soybean of the cry1 Gene from Bacillus thuringiensis

  • Received:2010-12-10 Revised:2011-01-04 Online:2011-04-05 Published:2011-04-05

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摘要：

此研究利用醋酸钠—抗生素加热法从土壤中分离获得1株产晶体的苏云金芽孢杆菌菌株（Bacillus thuringiensis）。通过已知cry1基因设计引物，以分离得到的菌株DNA为模板进行PCR扩增，将克隆到的目的片段进行测序。测序结果表明：该序列与cry1基因同源性达到90%~99%。将该基因连接到植物双元表达载体pBI121中，构建了该基因的植物表达载体，并将阳性重组质粒转化根癌农杆菌EHA105，利用农杆菌侵染子叶节的方法进行大豆的转化。通过初步的PCR验证，成功获得了转基因植株，建立了大豆的转化体系。

关键词: 柠条, 柠条, 药剂处理, 失水率, 苗木活力

Abstract:

A Bacillus thuringiensis strain which produced crystal was isolated from soil with the method of sodium acetate-antibiotics heating in this study. By the known cry1 gene sequences, a pair of primers was designed. The PCR amplification was completed with the template of the DNA of the isolated strain, and the cloned fragment was sequenced. The results of sequence analysis indicated that this fragment had at least 90%-99% homology compared with the other cry1 genes. The target fragment was ligated with pBI121 vector and the plant expression vector was constructed, which was transferred into the cotyledon node system in soybean by Agrobacterium tumefaciens-mediated method and severa1 transformed plants were obtained which tested by PCR amplification. This result indicated that the transformed system in soybean was established.

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