在家蝇中表达人胰岛素基因的研究  

Study on the Expression of Human Insulin Gene in Housefly

作　　者：李华[1] 刘维全[2] 刘海鹏[2] 王本旭[2] 王吉贵[3] 江禹[2] 

机构地区：[1]中国医科大学实验动物部,沈阳110001 [2]解放军军需大学军事兽医研究所,长春130062 [3]中国农业大学生物学院,北京100094

出　　处：《中国比较医学杂志》2003年第6期321-324,共4页Chinese Journal of Comparative Medicine

摘　　要：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BAEKON6 0 0 0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 10 μg 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 (F0 ) ,G4 18(3 0mg ml)筛选F1 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 2 0 7%、0 35 7%、0 4 73%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。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.

关 键 词：家蝇 胰岛素 基因表达 电穿孔 基因工程 

分 类 号：R977.15[医药卫生—药品]