家蚕bmnapi2基因的克隆与转录活性检测-中国农学通报.pdfVIP

中国农学通报 2011,27(01):351-355 Chinese Agricultural Science Bulletin 家蚕BmNaPi2基因的克隆与转录活性检测 杨金宏，孔卫青 （安康学院陕西省蚕桑重点实验室，陕西安康725000） 摘 要：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1336 bp （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1314 bp ，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。 关键词：家蚕；BmNaPi2 ；克隆；转录活性 中图分类号：S881.2 文献标志码：A 论文编号：2010-2129 CloningandTranscriptionalActivityDeterminationofSilkwormBmNaPi2Gene Yang Jinhong, Kong Weiqing (TheKeySericulturalLaboratoryofShaanxi, AnkangUniversity, Ankang Shaanxi 725000) Abstract: 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 3rd 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. Keywords:Bombyx mori; BmNaPi2; clone; transcriptional activity [6] 0 引言