家蚕核酸糖转运蛋白BmSlc35b3基因的克隆与序列分析

在电子克隆的基础上,利用RT-PCR方法克隆了家蚕的同源基因BmSlc35b3(GenBank登录号:GU244352)。生物信息学分析结果显示,该基因编码区长1128bp,包含5个外显子,由基因推定的蛋白质由375个氨基酸组成,内有9个跨膜结构域,8个N-糖基化位点,与家蚕溶质运载蛋白家族B3(ABD36159)有4个氨基酸的差异,与赤拟谷道(EFA00972)、金小蜂(XP_001601459)、致倦库蚊(EDS30138)、疟蚊(EAA11308)等的同源性均在70%以上。RT-PCR对BmSlc35b3基因在五龄3d家蚕幼虫9种组织中的表达进行检测,结果表明,其在中肠和精巢中表达,在其他组织中不表达。

Cloning and Sequence Analysis of the Nucleotide-sugar Transporter Gene BmSlc35b3 in Bombyx mori

The glycosylation by glycosyltransferases depends on the transport of nucleotide-sugar from the cytoplasm into the Golgi apparatus and the endoplasmic reticulum by NSTs.BmSlc35b3(GenBank accession number:GU244352),the homologous gene of silkworm,was cloned by RT-PCR.Bioinformatics analysis showed that this gene is 1128bp long,contains 5 exons and encodes 375 amino acids,including 9 transmembrane domains and 8 N-glycosylation sites.There are 4amino acids different from B3(ABD36159),one member of silkworm solute transporter family.Comparison of BmSlc35b3 to the homologues of Tribolium castaneum(EFA00972),Nasonia vitripennis(XP_001601459),Culex quinquefasciatus(EDS30138) and Anopheles gambiae(EAA11308) revealed above 70% similarity among their amino acid sequences.The results of RT-PCR showed that the genewas expressed only in midgut and testis among 9 tissues detected on the 3rd day of the fifth instar larvae.