羊BLG基因5′和3′调控区克隆及其调控GFP基因在乳腺细胞的表达  

Cloning of 5′, 3′ Flanking Sequence of Ovine BLG and Regulating the Expression of GFP in Mammary Gland Cell Line

作　　者：刘明军[1] 李文蓉[1] 武坚[1] 黄俊成[1] 郭志勤[1] 曲新勇[2] Paul Kroon 

机构地区：[1]新疆畜牧科学院,乌鲁木齐830000 [2]澳大利亚昆士兰大学生化系

出　　处：《生物工程学报》2002年第1期112-116,共5页Chinese Journal of Biotechnology

基　　金：国家 8 6 3高技术研究发展计划项目资助 (No .Z2 1 0 3 0 3)~~

摘　　要：and 3′ flanking region of ovine BLG were amplified from sheep genomic DNA according to the published whole sequence of ovine BLG and cloned to pGEM-T vector correspondently.By partially sequencing,the sequences of BLG 5′ and 3′ flanking were the same as that of publication completely.The recombinant structure used to direct exogenous gene especially to express in mammary gland was constructed by joining 4.2kb 5′ flanking with 2.1kb 3′ flanking.In order to assess the efficiency of BLG regulatory elements,green fluorescent protein (GFP) gene as a reporter was fused with BLG construct and transfected the mammary epithelial cells (TD47).Through observation under UV microscope and detection by fluorometer,it is demonstrated that the GFP has been successfully expressed in TD47 cell line.By virtue of direct observation and quantitative analysis,the BLG-GFP construct can be served as a model for the quick assessment of mammary gland expression construct.and 3′ flanking region of ovine BLG were amplified from sheep genomic DNA according to the published whole sequence of ovine BLG and cloned to pGEM-T vector correspondently.By partially sequencing,the sequences of BLG 5′ and 3′ flanking were the same as that of publication completely.The recombinant structure used to direct exogenous gene especially to express in mammary gland was constructed by joining 4.2kb 5′ flanking with 2.1kb 3′ flanking.In order to assess the efficiency of BLG regulatory elements,green fluorescent protein (GFP) gene as a reporter was fused with BLG construct and transfected the mammary epithelial cells (TD47).Through observation under UV microscope and detection by fluorometer,it is demonstrated that the GFP has been successfully expressed in TD47 cell line.By virtue of direct observation and quantitative analysis,the BLG-GFP construct can be served as a model for the quick assessment of mammary gland expression construct.

关 键 词：羊BLG基因 3′调控区 调控 GFP基因 羊乳球蛋白 基因克隆 绿色荧光蛋白 乳腺细胞系 表达 

分 类 号：Q786[生物学—分子生物学]