牛β-乳球蛋白基因的克隆及其调控外源基因表达的研究  

The Cloning of Bovine Beta-lactoglobulin Gene and the Identification of Its Effect for Regulating the Expression of Foreign Gene

作　　者：陈红星[1] 程萱[1] 杨晓[1] 邓继先[1] 苏国富[1] 黄培堂[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100071

出　　处：《中国实验动物学报》2001年第2期78-82,共5页Acta Laboratorium Animalis Scientia Sinica

基　　金：国家863计划! (Z2 1 0 3 0 1 )

摘　　要：目的　制备乳腺生物反应器所必需的乳腺特异性表达的调控序列 ,并验证其指导外源基因表达的能力。方法　用PCR法从奶牛染色体上分 5段扩增出了全长 8 2Kb的牛 β 乳球蛋白基因 ,包括 1 8Kb的 5′侧翼区、1 7Kb的 3′侧翼区及 4 7Kb的gDNA区。扩增出的各片段克隆到T 载体上 ,酶切鉴定及序列分析均证实了所扩增片段的正确性。将五个片段与荧光素酶cDNA拼接成荧光素酶瞬时表达载体并在小鼠乳腺中瞬时表达。结果　注射荧光素酶瞬时表达载体的小鼠乳汁中明显测出了荧光素酶活性。结论　所克隆的牛 β 乳球蛋白基因表达调控序列能够指导外源基因在小鼠乳腺中表达。Objective\ To get the regulatory elements which are essential for generating mammary gland bioreactors,and identify their effect for regulating the expression of foreign gene.Methods\ The whole 8.2 Kb bovine beta-lactoglobulin gene was amplified by PCR method,including the 1.8 Kb 5′ flanking region,4.7 Kb gDNA region,1.7 Kb 3′ flanking region.All the amplified fragments were cloned in T-vectors and were proved to be correct by restrction enzyme digestion and sequencing analysis.The five fragments and the luciferase cDNA were ligated together to get the luciferase temporary expression vector,which was temporarily expressed in the mammary gland of mice.Results\ The luciferase activity was detected in the milk of mice whose mammary glands were injected with the expression vector.Conclusion\ The cloned bovine BLG gene could direct foreign gene expression in the mammary gland of mice.\;

关 键 词：小鼠 乳腺 克隆 扩增 荧光素酶 瞬时表达 载体 外源基因表达 Β-乳球蛋白基因 表达调控 

分 类 号：Q78[生物学—分子生物学] Q819