家蚕BmNaPi2基因的克隆与转录活性检测

Cloning and Transcriptional Activity Determination of Silkworm BmNaPi2 Gene

通过生物信息学分析和分子生物学实验获得了家蚕钠离子依赖性的磷酸盐转运蛋白的同源基因BmNaPi2。该基因长1336bp(GenBank登录号:HM593516),含5个外显子,位于家蚕第3号染色体,编码区长1314bp,基因编码437个氨基酸,预测蛋白序列有10个疏水的跨膜结构域,与果蝇、疟蚊、埃及伊蚊、致倦库蚊、赤拟谷道等的同源蛋白的相似性约57%。RT-PCR检测基因在5龄第3天家蚕幼虫9种组织中的血液、马氏管和体壁中表达,而中肠、丝腺、生殖腺、头和脂肪体中没有表达。

The homologous gene of Na （＋）-dependent phosphate cotransporter （NaPi） in silkworm, BmNaPi2, was obtained through bioinfomatics analysis and molecular biological experiment. The gene was located on 3 rd chromosome, 1336 bp long （GenBank accession numer： HM593516）, and 5 exons inside. The ORF of gene was 1314 bp in length and encodes 437 amino acids, with 10 hydrophobic transmembrane domains. A comparison of BmNaPi2 with homologous protein from Drosophila melanogaster, Anopheles gambiae, Aedes aegypti, Culex quinquefasciatus, Tribolium castaneum revealed that their amino acids share about 57% similarity. It is shown by RT-PCR experiment that the BmNaPi2 had expression in blood, malpighian tubule and integument, while no expression in midgut, silk gland, gonad, head, and fat body among 9 tissues detected on day 3 of the fifth instar larvae.