重组汉坦病毒包膜糖蛋白G1、G2基因体内外表达的评价  

Evaluation on Expression of Hantavirus Z37 Envelope Glycoprotein Genes G1 and G2 Recombinants in Vitro and in Vivo

作　　者：黄玉仙[1] 翁心华[1] 瞿涤[2] 朱函坪[3] 陆群英[3] 朱智勇[3] 

机构地区：[1]复旦大学附属华山医院传染科,上海200040 [2]上海医学院医学分子病毒实验室,上海200032 [3]浙江省疾病控制中心肾综合征出血热实验室,杭州310009

出　　处：《复旦学报（医学版）》2004年第6期603-606,共4页Fudan University Journal of Medical Sciences

摘　　要：目的　构建分别编码汉坦病毒浙 37(Z37)株包膜糖蛋白G1、G2的真核表达质粒 ,并在离体及活体内评价重组汉坦病毒 (HV)包膜糖蛋白G1、G2基因的表达。方法　将编码G1、G2的基因片段分别插入至真核表达载体 pcDNA3.1(+) ,获重组质粒pcDNA3.1 G1、pcDNA3.1 G2。在体外转录翻译系统和真核细胞中表达重组基因 ,并在BALB/C小鼠中评价包膜糖蛋白G1、G2所激发的特异性体液免疫应答效果。结果　重组质粒pcDNA3.1 G1、pcDNA3.1 G2转染COS 7细胞后 ,IFA法可检测到细胞内有特异性荧光分布 ;在体外蛋白质转录、翻译系统 ,重组质粒 pcDNA3.1 G1、pcDNA3.1 G2所表达的蛋白质分子量分别约为 70KD及 5 5KD ;在免疫的部分小鼠体内可检测到特异性抗体。结论　成功地构建了分别编码HVZ37株包膜糖蛋白G1、G2的重组质粒 。Purpose: To construct Hantavirus Z37 envelope glycoprotein genes G1 and G2 recombinants and evaluate the expression of the recombinants in vitro and in vivo. Methods: The clone fragment coding G1input commas G2 envelope glycoprotein were inserted into a eukaryote expression vector pcDNA3.1 respectively to construct the recombinant plasmids pcDNA3.1-G1 and pcDNA3.1-G2. The recombinant plasmids were expressed respectively in a coupled transcription/ translation system in vitro and in transfected COS-7 cells, and the efficiency of recombinant plasmids in eliciting humoral immune response was evaluated in BALB/C mice. Results: After transfection of the recombinant expression plasmids pcDNA3.1-G1and pcDNA3.1-G2, specific antigen were detected within cells by IFA. HV Z37 envelope glycoprotein G1 and G2 molecular weight were about 70KD and 55KD respectively. Serum anti-hantavirus antibodies were detected in BALB/C mice immunized with the recombinant plasmids. Conclusions: The reombinant expression plasmids pcDNA3.1-G1input commas pcDNA3.1-G2 coding HV Z37 envelope glycoprotein G1 and G2 respectively were constructed and expressed in vitro and in vivo successfully.

关 键 词：包膜糖蛋白 体内 表达 PCDNA3 汉坦病毒 重组质粒 基因 蛋白质分子 转录 真核细胞 

分 类 号：R512[医药卫生—内科学] R373[医药卫生—临床医学]