GbTCP10基因植物表达载体的构建及拟南芥转化  

Construction of Plant Expression Vector of Gb TCP10 Gene and Transformation of Arabidopsis thaliana

作　　者：王怡[1] 于月华[1] 杨成元 陈全家[1] 倪志勇[1] Wang Yi;Yu Yuehua;Yang Chengyuan;Chen Quanjia;Ni Zhiyong(College of Agriculture,Xinjiang Agricultural University,Urumqi,830052)

机构地区：[1]新疆农业大学农学院

出　　处：《分子植物育种》2020年第1期144-149,共6页Molecular Plant Breeding

基　　金：国家自然科学基金项目(U1703115);南农-新农联合基金项目(KYYJ201801);天山青年计划项目(2018Q002;2018Q018)共同资助

摘　　要：TCP转录因子广泛参与植物细胞生长和增殖调控,本研究构建海岛棉GbTCP10基因沉默和过量表达植物表达载体。以GbTCP10基因pGEM-T Easy-GbTCP10质粒为模板进行PCR扩增,连接至植物表达载体pCAMBIA3301中,再利用冻融法和热激法转到农杆菌GV3101菌株中,通过农杆菌浸染法转化拟南芥植株,初步证明已获得海岛棉GbTCP10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amiRTCP10前体的植物表达载体amiRTCP10-pCAMBIA3301,本试验结果有助于进一步研究海岛棉GbTCP10基因的生物学功能和棉花纤维发育机制。TCP transcription factor is widely involved in plant cell growth and proliferation regulation.GbTCP10 gene silencing and overexpression plant expression vector were constructed in this study.The GbTCP10 gene pGEM-T Easy-GbTCP10 plasmid was used as the template for PCR amplification.The gene was constructed into the plant expression vector p CAMBIA3301,transformed into Agrobacterium tumefaciens strain GV3101 by freeze-thaw method,and transformed into Arabidopsis plants by Agrobacterium tumefaciens method.It was initially proved that GbTCP10 gene of Arabidopsis thaliana was obtained.To construct a plant expression vector amiRTCP10-p CAMBIA3301 containing the amiRTCP10 precursor by overlapping PCR method for replacing mature Arabidopsis thaliana At-MIR319,so as to lay the foundation for further study on the biological function of GbTCP10 gene and provide a theoretical basis for study development mechanism of cotton fiber.

关 键 词：海岛棉(Gossypium barbadense) 拟南芥(Arabidopsis thaliana) 植物表达载体 amiRNA 

分 类 号：Q943.2[生物学—植物学]