含增强型绿色荧光蛋白(EGFP)基因的家蚕同源重组质粒载体的构建  

Construction of a Homologous Recombination Plasmid Vector pMD-Fib-IE-EGFP Carrying EGFP as Reportor Gene

作　　者：桂慕燕[1] 叶向群[1] 吴春旭[1] 熊秀芳[1] 

机构地区：[1]厦门大学生命科学学院,细胞生物学与肿瘤细胞工程教育部重点实验室,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2002年第5期628-633,共6页Journal of Xiamen University：Natural Science

基　　金：国家自然科学基金 (39870 410 );厦门大学校级自选课题 (Y0 70 14)资助项目

摘　　要：以含家蚕丝心蛋白重链基因同源片段的质粒pG35 0为出发质粒 ,插入增强型绿色荧光蛋白 (EGFP)基因 ,构建成以EGFP为报告基因的同源重组质粒载体 pMD Fib IE EGFP ,通过PCR及酶切鉴定表明EGFP以正确的方式插入到原始质粒中 ,通过脂质体介导法转染胃癌细胞能发出很强的绿色荧光 ,表明该质粒能够在真核细胞中表达 。A new plasmid vector pMD-Fib-IE-EGFP which contains enhanced green fluorescent protein gene as reportor gene was constructed by subtitute EGFP for GFP in the original plasmid pG350. The recombinant plasmid was identified by restriction endonuclease enzyme analysis and PCR. The recombined plasmid was transferred to stomach cancer cell mediated by lipofectine. The transfected cell expressed strong fluorescence. The constructed plasmid vector provided foreign DNA material for transgenic silkworm.

关 键 词：EGFP 家蚕 同源重组质粒载体 增强型绿色荧光蛋白基因 转基因蚕 载体构建 基因表达 

分 类 号：S881.2[农业科学—特种经济动物饲养] Q782[农业科学—畜牧兽医]