在家蝇中表达人胰岛素基因的研究

目的 研究人胰岛素基因在家蝇中的表达，建立家蝇生物反应器。方法 以家蝇作为实验动物，利用BAEKON6000型基因转移仪将胰岛素基因组基因导入家蝇进行实验研究。收集家蝇新产卵(1—1．5h内)，将环状质粒pCMV—mINS、pCMV-ImINS、pEF1α—mINS、pEF1α—ImINS以10μg/ml浓度分别与新采集的家蝇卵混合后进行电转移。经人工孵化发育为成虫(R)，G418(3．0mg／ml)筛选Fl代，待F1代发育至成虫并产卵后，采用PCR、Tricine-SDS-PAGE和Western-blot技术对F2代蝇蛆进行整合与表达水平检测。结果 四种质粒均能在家蝇中表达人胰岛素，表达量各占蝇蛆总蛋白0．207％、0．357％、0．473％和0．831％。结论 人胰岛素基因能够在蝇蛆中表达，即蝇蛆机体可对人胰岛素原进行翻译后加工。

Study on the Expression of Human Insulin Gene in Housefly

Objective To construct bioreactor with housefly to express human insulin.Methods Four recombinant expression plasmids were transferred respectively into the new fertillized eggs of housefly by electroporation.The first generation was screened with G418 after the founders were incubated to adults.Human insulin was detected by radioimmumoassay, Tricine-SDS-PAGE and Western-blot in samples of maggot of the second generation collected at the time of 96?hours.Results All four recombinant expression plasmids could express human insulin in housefly. Conclusion The housefly maggot has the ability of processing post-translation insulin genomic gene.So housefly can be used as bioreactor to produce human insulin.