家蚕Bmsps1基因的克隆和原核表达

Cloning and Expression of Bmsps1 of Bombyx mori

从家蚕EST数据中检索到果蝇(D.melanogaster)硒磷酸合成酶基因(patuf)的氨基酸序列,用seq MAN延伸,电子克隆家蚕sps1的cds,用PCR克隆家蚕sps1基因序列。家蚕sps1基因cDNA长1817bp(登录号:ABA43639.1)。利用BLASTN进行同源性分析表明与果蝇的同源性为94%,与大肠杆菌序列同源性最差为46%。Bmsps1基因在家蚕基因组中是单拷贝的,只有一个外显子。推导3'UTR序列包含AATAA终止信号和Poly(A)。同时我们对Bmsps1进行了原核表达研究。

The amino acid sequence of selenophosphate synthetase 1 （SPS1） from Drasophila melanogaster was subjected to tblastn searching EST of Bombyx mori, and cds of Bmspsl of B. mori was electronically cloned. The cDNA of selenophosphate synthetase 1 of B. mori is 1817 bp （accession ABA43639.1）. Blastn analysis revealed high similarity （94%） between Bmspsl gene and D. melanogaster sps 1 gene and low similarity （46 %） between Bmspsl gene and E. coli spsl gene. Blastn analysis with genome data revealed that bmgapdh is a single copy located on the genome and has only one exon. 3＇UTR is speculated to contain putative stop codon （AATAA） and Poly（A）. Prokaryotic expression of Bmspsl in E. coli was studied.