家蚕Bm050524基因的表达、纯化及转录分析

he gene Bm050524 is a novel gene from silkworm, Bombyx mori . Up to now, there is no literature reporting about any characters about this gene. Bm050524 contains an open reading frame(ORF) of 483bp nucleotides , and encodes 160 amino acids with the predicted molecular weight of 18.56 kDa. We also took genome alignment between Bm050524 cDNA and silkworm genome, and the results showed that this gene includes three exons. The recombinant protein of His-Bm050524 was expressed in E.coli BL21 (DE3) and existed as inclusion body. After purification by nickel affinity column chromatography, polyclonal antibody was obtained. Total protein of seven tissues of the fifth instar larvae were extracted and analyzed by Fluorescent Western blotting. The results showed that Bm050524 widely existed in the tissues of the fifth instar larvae. Total RNA of nine tissues of the fifth instar larvae (Bombyx.mori) were extracted for RT-PCR experiment. RT-PCR demonstrated there were high transcriptional level head compared to other tissues. These results laid a good foundation of further studies of Bm050524.