人α－干扰素基因在小菜蛾细胞中的表达及其影响因素探讨  

EXPRESSION OF HUMAN α INTERFERON GENE IN PLUTELLA XYLOSTELLA CELLS

作　　者：马春红[1] 施先宗[1] 余泽华[1] 杨凯[1] 陈曲侯[1] 

机构地区：[1]华中师范大学昆虫学研究所,山东医科大学

出　　处：《华中师范大学学报（自然科学版）》1996年第2期212-215,共4页Journal of Central China Normal University：Natural Sciences

摘　　要：将含有人α－干扰素（ＩＦＮα）基因的杆状病毒转移载体（ｐＡｃ３７３－ＩＦＮα）与苜蓿银纹夜蛾核型多角体病毒（ＡｃＮＰＶ）ＤＮＡ经钙磷沉淀法共转染小菜蛾细胞系（ＢＣＩＲＬ－Ｐｘ２－ＨＮＵ３，Ｐｘ），空斑纯化后，利用细胞致病效应（ＣＰＥ）抑制法检测得到了干扰素效价（６８７２．６ＩＵ），证明得到了可以在Ｐｘ细胞中表达ＩＦＮα的重组病毒（Ａｃ－ＩＦＮα）．并探讨了培养温度和血清浓度对ＩＦＮα表达水平的影响，结果表明：在２８℃时，培养基中补加２％的小牛血清，可获得最高的ＩＦＮα效价（１７４３５．９ＩＵ）．In this paper, Human α interferon(IFNα) gene carried by a baculovirus transfer vector (pAc373-IFNα) was transferred into wild AcNPV genome by the calcium phosphate co-precipitation in Plutella xylostella cells (Px cells ). A polyhedrin negative(occ-) recombinant virus Ac-IFNα was obtained by plaque assays. The Expression of IFNα was determined by minor cytopathic effect(CPE) reduction assay. The titer of IFNα in the media was 6 872. 6 IU,Moverover, the effects of temperature and serum concertration on the expression level of IFNα were also studied here. Results showed that 28℃ and lower serum concretration(2%) were more suitable for the high expression of IFNα.

关 键 词：α-干扰素基因 杆状病毒 基因表达 小菜蛾细胞系 

分 类 号：Q782[生物学—分子生物学]