GhMYB4基因过量表达载体的构建及拟南芥转化

Construction of GhMYB4 Gene Over-expression Vectors and Transformation of Arabidopsis thaliana

作为广泛参与植物体内的多种生理生化反应的MYB转录因子是植物中最大的转录因子家族之一,为阐明GhMYB4基因在棉纤维发育过程中的功能,本研究构建GhMYB4基因过量表达载体,以pGEM-T Easy-GhMYB4质粒为模板进行PCR扩增,成功将该基因构建到植物表达载体pEGAD中,并利用冻融法转入农杆菌GV3101菌株,通过农杆菌浸染法转化拟南芥植株,经过含草甘膦成分的除草剂筛选和PCR检测初步证明已获得转GhMYB4基因的拟南芥。为后期探讨转化棉花GhMYB4基因的生物学功能和棉花纤维发育机制提供理论参考。

MYB transcription factors, which are involved in many physiological and biochemical reactions in plants, are one of the largest transcription factors in plants. In order to elucidate the function of GhMYB4 gene in cotton fiber development. In this study, the overexpression vector of GhMYB4 gene was constructed using pGEM-T Easy-GhMYB4 plasmid as template for PCR amplification. The gene was successfully constructed into the plant expression vector p EGAD. Then, it was transformed into Agrobacterium tumefaciens GV3101 strain by freezing and thawing method, and transformed into Arabidopsis thaliana plant by Agrobacterium tumefaciens soaking method. After herbicide containing glyphosate screening and PCR detection, it was preliminarily proved that the Arabidopsis thaliana transformed with GhMYB4 was obtained. It would provide a theoretical reference for the later study on the biological function and cotton fiber development mechanism of transformed cotton GhMYB4 gene.