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中国农学通报

中国农学通报 ›› 2007, Vol. 23 ›› Issue (4): 82-82.

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拟南芥热激启动子AtHSP70b的克隆与功能分析

裴华丽,胡华刚,张兴国,苏承刚,宋孝霞   

  • 出版日期:2007-04-05 发布日期:2007-04-05

Cloning and Functional Analysis of the Heat-inducible Promoter AtHSP70b

Pei Huali, Hu Huagang, Zhang Xingguo, Su Chenggang, Song Xiaoxia   

  • Online:2007-04-05 Published:2007-04-05

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摘要： 从拟南芥Columbia生态型(Arabidopsis thaliana)的基因组DNA中扩增出热激启动子AtHSP70b基因，并检测其热激表达的严谨性。将热激启动子AtHSP70b插入到pSAU2008的gus基因及NOS终止子上游，构成热激启动子控制GUS报告基因的表达盒“AtHSP70b-GUS-Tnos”，并将其插入到pVCT2020中得到表达载体pV-HSP70bGUS。通过冻融法将其导入根癌农杆菌（Agrobacterium tumefaciens）LBA4404，并转化烟草(W38)获得再生植株，通过PCR检测鉴定，表明“AtHSP70b-GUS-Tnos”表达盒已整合到烟草基因组中。并转化烟草检测启动子的表达活性。序列分析表明，克隆的启动子长度204bp与目前已经发表的启动子序列完全一致。GUS组织化学法检测证明启动子AtHSP70b在烟草中具有高温诱导表达的能力。但在常温下也有不同程度的表达，因此需要对该启动子进行序列改造以增强其热激表达的严谨性。

关键词: 豆类丝核菌, 豆类丝核菌, 苦马豆素, 次级代谢产物, 肿瘤, H22

Abstract: The promoter of a heat-inducible gene AtHSP70b was amplified from the genome DNA of Arabidopsis thaliana cv. Columbia, which had a length of 204bp and the same sequence as published previously. The expression vectors, pV-HSP70bGUS was constructed by inserting the promoter of AtHSP70b. The vector was introduced into Agrobacterium tumefaciens LBA4404 which was used to mediate the transfection of AtHSP70b promoter fragment into the tobacco (W38). The regenerated plants were analyzed and showed that the promoter fragment was integrated into the genome of tobacco. Histochemical staining of GUS activity of the transgenic plants proved that the promoter of AtHSP70b expressed unprecisely.

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