与ATP6相互作用的蛋白质的筛选和Bn HDL基因的cDNA克隆

Screening of proteins interacting with ATP6 and the cDNA cloning of the Bn HDL gene

以与甘蓝型油菜波里马细胞质雄性不育有关的atp6基因作为诱饵蛋白,通过酵母双杂交技术,筛选甘蓝型油菜的cDNA文库,寻找与其相互作用的蛋白质。根据筛选所获得的其中一个阳性克隆的序列,利用RACE技术和TAIL PCR技术,获得该基因的全长cDNA序列。将氨基酸序列在NCBI上进行Blast比对分析,具有水解酶家族17的保守结构域,与拟南芥的beta-1,3-glucanase 5具有84%的同源性,可能为水解酶家族的一个成员,暂定该蛋白质为Bn HDL。

The atp6 gene that correlated with Brassica napus Pol cytoplasmic male sterility was used as the bait protein, by using yeast two-hybrid system, the Brassica napus cDNA library were screened, identified the proteins interacting with ATP6. According to the sequence of a positive clone, the fulllength cDNA sequence was obtained by RACE method and TAIL PCR method. Based on the sequence of the amino acid, BLAST analysis against NCBI database was performed. The BLAST results indicate that putative conserved domains（Glyco_ hydro_17） have been detected and have 84% identities with beta-1,3 -glucanase 5 （Arabidopsis thaliana）. It was presumed that maybe a member of hydrolase family and was named Bn HDL gene temporarily.