海岛棉GbTCP5基因沉默和过量表达载体的构建及拟南芥转化
Construction of GbTCP5 Gene Silencing and Over-expression Vectors and Transformation of Arabidopsis

海岛棉具有优良的纤维品质，TCP蛋白参与细胞生长和增殖调控，本研究构建海岛棉GbTCP5基因沉默和过量表达植物表达载体。以GbTCP5基因pGEM-T Easy-GbTCP5质粒为模板进行PCR扩增，并将该基因构建到植物表达载体pCAMBIA3301中，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，初步证明已获得转化海岛棉GbTCP5基因的拟南芥。利用重叠PCR方法替换拟南芥 At-MIR319的成熟链和互补链序列，构建含有 amiRTCP5前体的植物表达载体amiRTCP5-pCAMBIA3301，为深入研究海岛棉GbTCP5基因的生物学功能奠定基础，为研究棉花纤维发育机制提供理论依据。
Gossypium barbadense has the best cotton fiber and TCP protein is involved in the regulation of cell growth and proliferation. GbTCP5 gene silencing and over-expression vector were constructed in this study. The GbTCP5 gene pGEM-T Easy-GbTCP5 plasmid were used as a template for PCR amplification. The gene was constructed into the plant expression vector pCAMBIA3301, transformed into Agrobacterium tumefaciens strain GV3101 by freeze-thaw method, and transformed into Arabidopsis plants by Agrobacterium tumefaciens method. It was initially proved that GbTCP5 gene of Arabidopsis thaliana was obtained.To construct a plant expression vector amiRTCP5-pCAMBIA3301 containing theamiRTCP5 precursor by overlapping PCR method for replacing mature Arabidopsis thaliana At-MIR319,so as tolay the foundation for further study on the biological function of GbTCP5 gene and providing a theoretical basis for studying development mechanism of cotton fiber.