摘要： 磷酸盐是蚕必需的无机物,钠离子依赖性的磷酸盐转运蛋白(Na(+)-dependent phosphate cotransporter,NaPi)调节磷酸盐在生物体内的代谢和转运.本研究通过生物信息学分析和分子生物学试验获得了家蚕NaPi的同源基因BmNaPi,该基因位于家蚕8号染色体,编码区长1 437 bp,有10个外显子,编码478个氨基酸,预测蛋白序列在氨基末端含有一信号肽,序列中间有8个转膜结构域,与登录号为AAF7968、XP_001602874、EAA00281、XP-39379、XP00190194等同源蛋白的一致性均在70%及以上.RT-PCR检测该基因在龄3 d家蚕幼虫9种组织中的血液、体壁、生殖腺和头中表达,而中肠、丝腺、马氏管和脂肪体中没有表达.最后成功构建了该基因的酵母穿梭表达质粒pGBKT7-BmNaPi.

关键词: 家蚕, BmNaPi, 克隆, 组织表达, 酵母穿梭表达质粒

Abstract: Na(+)-dependent phosphate cotransporter(NaPi) mediate the transport and metabolism of inorganic phosphate.Phosphate is essential for silkworm and the homologous gene of silkworm,BmNaPi,is obtained through bioinfomatics analysis and molecular biological experiment.The gene is located on 8th chromosome,1 437 bp long,and encodes 478 amino acids,with a signal peptide in the N-termi and 8 transmembrane domains inside.A comparison of BmNaPi with homologous protein AAF57968,XP_001602874,EAA00281,XP_393759,XP_001950194 from other sources revealed that their amino acids share above 70% identity.It is shown by RT-PCR experiment that the BmNaPi has expression in blood,integument,gonad and head,while no expression in midgut,silk gland,malpighian tubule and fat body among 9 tissues detected on day 3 of the fifth instar larvae.Then yeast shuttling expression plasmid pGBKT7-BmNaPi was successfully constructed.

Key words: Bombyx mori, BmNaPi, Clone, Tissue expression, Yeast shuttling expression plasmid

中图分类号: 

• S881.2