棉花HSP70基因植物表达载体的构建及转化烟草的研究

为研究棉花HSP70 基因在转基因植物中抗旱效果，利用前期克隆的GhHSP70 基因，以pCAMBIA1304 质粒为植物表达载体，构建了GhHSP70 基因的植物表达载体CAMBIA1304-HSP70；采用冻融法转入根癌农杆菌EHA105 菌株，通过叶盘法对模式植物烟草进行遗传转化研究。结果表明，在潮霉素(50 mg/L)选择压力下获得的烟草转化不定芽和完整植株，经过PCR方法以及GUS基因组织化学法检测鉴定转基因烟草，其中有5 株为阳性植株。初步证实了GhHSP70 基因已导入烟草基因组中。为进一步研究该基因的功能提供实验基础，同时为后续转化棉花改善其抗旱能力奠定基础。

Research on Plant Expression Vector Construction and Transformation to Nicotiana tabacum of Cotton HSP70 Gene

In order to study the response of cotton HSP70 gene to drought stress in transgenic plants, the pCAMBIA1304-HSP70, plant expression vector, was constructed by using the cloned GhHSP70 gene and the pCAMBIA1304 vector. The pCAMBIA1304- HSP70 was transformed into EHA105 strain by freeze- thaw method. Then, the genetic transformation of tobacco was studied through leaf dish transformation method. Results showed that the tobacco adventitious buds and complete plants were obtained under Hygromycin (50 mg/L) selective pressure, the transgenic tobaccos were detected by GUS gene histochemical method and PCR method, and five of them were positive plants. We preliminarily confirmed that the GhHSP70 gene had transferred into tobacco genome. This study provided a fundamental experiment condition for further research on the GhHSP70 gene, and it might lay the foundation for genetic transformation in order to improve the drought resistance of cotton.