人生长激素基因在昆虫细胞中表达的初步研究  

STUDY ON THE EXPRESSION OF HUMAN GROWTH HORMONE GENE IN INSECT CELLS

作　　者：俞新大[1] 耿朝晖[1] 周卫东[1] 张宝珠[1] 张爱虹 李建民[1] 沈孝宙[2] 王瑛[2] 

机构地区：[1]南开大学生命科学学院,天津300071 [2]中国科学院北京动物研究所,北京100080

出　　处：《南开大学学报（自然科学版）》1998年第3期98-101,共4页Acta Scientiarum Naturalium Universitatis Nankaiensis

基　　金：国家"九五"重点科技攻关项目

摘　　要：将人生长激素hGH的cDNA片段克隆在转移载体pVL1393的多角体蛋白基因启动子下游，构成重组转移载体pVL1393hGH，将该质粒DNA与昆虫杆状病毒（AcNPV）DNA共转染秋粘虫（Spodopterafrugiperda）细胞Sf9，通过体内同源重组，hGH基因取代多角体蛋白基因，从而整合到病毒基因组上，经感染昆虫细胞后表达外源hGH基因，通过反复病毒空斑纯化，获不含多角体的重组病毒，利用免疫化学发光法测定hGH表达量达40μg／mL．The recombinant transfer vector pVL1393hGH has been constructed in which the human growth hormone cDNA was inserted under the control of the polyhedron gene promoter. The Spodoptera frugiperda (Sf9) cells were co-trans fected with both the plasmids containing hGH cDNA and wild-type AcNPV DNA.The hGH cDNA is transferred to the AcNPV genome by homologous recombination,and the recombinant virus R-AcVhGH was obtained by multiple plaque purification. The high level of production of hGH (40μg/mL)in supernatant of the infected monolayer culture and suspension culture was determined by immunochemiluminescent assay.

关 键 词：hGH基因 ACNPV 转移载体 昆虫细胞 表达 

分 类 号：Q78[生物学—分子生物学] Q753