拟南芥AtSb10基因的克隆及其过表达转基因株系的获得

Cloning of AtSb10 gene in Arabidopsis thaliana and over-expression of AtSb10 in Arabidopsis thaliana

拟南芥At Sb10基因是一个表达谱和功能都未知的基因,生物信息学分析发现其氨基酸序列含有2个连续的MATH结构域。为分析At Sb10基因的功能,首先,利用RT-PCR方法检测At Sb10基因在拟南芥中的表达模式,结果表明,该基因只在根中表达,而茎、叶片、花与果荚中未检测到;然后克隆了At Sb10基因全长,构建了过量表达载体,通过农杆菌介导法将该载体转入拟南芥Col-0中,获得了10株过表达转基因植株,为进一步研究At Sb10基因的功能奠定了基础。

Bioinformatic analysis suggested that At Sb10,whose function and expression profile were unknown,contained two consecutive MATH domain structures. In order to further explore the function of At Sb10 gene,RT-PCR was used to investigate the expression profile of Atsb10 in Arabidopsis thaliana. The results showed At Sb10 gene was expressed only in roots,not in stems,leaves,flowers and siliques. Meanwhile,full-length of At Sb10 was cloned into overexpression binary vector p CAMBIA1300 driven by the Ca MV35 S promoter. Then this expression vector was transformed into Columbia-0 ecotype mediated by Agrobacterium tumefaciens GV3101. Ten over-expression transgenic plants were obtained,which would provide good materials for further studying on the function of Atsb10.