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中国农学通报

中国农学通报 ›› 2012, Vol. 28 ›› Issue (27): 189-193.

所属专题： 生物技术

• 生物技术科学 • 上一篇    下一篇

INO启动子克隆及胚珠特异表达载体的构建

刘娜 刘兴地 李明芳 郑学勤   

  • 收稿日期:2011-12-28 修回日期:2012-02-27 出版日期:2012-09-25 发布日期:2012-09-25
  • 基金资助:

    中央级公益性科研院所基本科研业务费专项资金资助项目

Cloning of INO promoter and construction of ovule-specific expression vector

  • Received:2011-12-28 Revised:2012-02-27 Online:2012-09-25 Published:2012-09-25

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摘要：

为了观察转基因植株胚珠特异性表达，采用CTAB法提取拟南芥基因组DNA，构建了35S::pCAMBIA1304-L117载体并对番茄进行遗传转化。根据已报道的INO启动子序列设计并合成一对特异引物，通过PCR对启动子进行扩增、连接并对其进行测序，结果表明：与GenBank中已注册的INO启动子序列的同源性为100%，并且构建了以INO代替CAMV35S启动子的pCAMBIA1304-L117植物表达载体，采用冷冻法将其转入根癌农杆菌EHA105。成功构建了35S::pCAMBIA1304-L117载体，并为进一步检测奠定了基础。

关键词: 调查, 调查

Abstract:

The purpose of this study was to study the ovule-specific expression of the transgenic plants. Genomic DNA was extracted from Arabidopsis by using CTAB method, 35S::pCAMBIA1304-L117 gene plant expression vector was contructed, and then the genetic transformation of tomato was carried out. According to the reported INO promoter sequence, a pair of special primers was designed and synthesized. They were used to amplify by PCR, and this fragment was inserted into vector and sequenced. The results showed the cloned fragment had a homology of 100% with the INO promoter logged on GenBank. Furthermore the plant expression vector of pCAMBIA1304-L117 replacing the CAMV35S promoter with INO was constructed and transformed into agrobacterium EHA105 by the freezing method.The 35S::pCAMBIA1304-L117 gene plant expression vector was constructed,and it will lay a more foundation.