家蚕磷酸吡哆醇氧化酶基因的表达谱分析  

Expression profiling of pyridoxine 5′-phosphate oxidase gene in Bombyx mori

作　　者：葛俊楠[1] 张剑韵[2] 黄龙全[1] 

机构地区：[1]安徽农业大学茶与食品科技学院,合肥230036 [2]安徽农业大学生命科学学院,合肥230036

出　　处：《昆虫学报》2011年第8期877-880,共4页Acta Entomologica Sinica

基　　金：国家自然科学基金项目(30870338)

摘　　要：【目的】了解家蚕Bombyx mori维生素B6关键代谢酶磷酸吡哆醇氧化酶(pyridoxine-5'-phosphate oxidase,PNPO)基因在家蚕不同发育阶段及5龄幼虫不同组织中的表达差异。【方法】将家蚕PNPO基因的重组表达质粒pET-22b(+)-PNPO转化入大肠杆菌Escherichia coli Rosetta中诱导表达,纯化蛋白制备多克隆抗体。分别采用荧光定量PCR和Western blot方法对家蚕PNPO基因进行了转录水平和翻译水平的表达分析。【结果】在家蚕发育水平上,5龄幼虫的PNPO翻译量为最高。PNPO基因在5龄幼虫各组织中的转录水平由高到低依次为精巢、头、中肠、马氏管、卵巢、表皮、脂肪体、丝腺;翻译量也以精巢为最高,其次是头、中肠和马氏管。【结论】明确了PNPO在家蚕各发育阶段及5龄幼虫各组织中的表达情况。[Aim] The expression profile of gene encoding pyridoxine phosphate oxidase（PNPO） ,which is a key enzyme related to VB6 metabolism,was analyzed in different developmental stages and tissues of the 5th instar larvae of Bombyx mori. [Methods] The recombinant plasmid pET-22b（＋） -PNPO was transformed to Escherichia coli Rosetta for induction,expression and purification of PNPO,and then the polyclonal antibody was prepared. The expression profile of PNPO gene was analyzed by Real-time PCR and Western blot,respectively. [Results] The highest translation level of PNPO gene was found in the 5th instar larvae. The transcription level in tissues of 5th instar larvae was in sequence of testisheadmidgutMalpighian tubulesovarycuticlefat bodysilk glands. However,the highest translation level was found in testis,and then was in head,midgut,and malpighian tubules,respectively. [Conclusion] The expression profile PNPO gene in B. mori has been defined by this study.

关 键 词：家蚕 磷酸吡哆醇氧化酶 WESTERN BLOT 荧光定量PCR 表达谱 

分 类 号：Q966[生物学—昆虫学]