小鼠乳清酸蛋白上游调控序列的克隆及其指导下CAT基因在家兔乳腺中的表达  

Amplification of the 5′Upstream Region of Mouse Whey Acid Protein Gene and its Guidance of the Expression of CAT Gene in Rabbit Mammary Gland

作　　者：杜卫华[1] 赵君[1] 张守峰[1] 任文陟[2] 扈荣良[1] 

机构地区：[1]解放军军需大学军事兽医研究所,吉林长春130062 [2]解放军军需大学军事兽医系,吉林长春130062

出　　处：《兽医大学学报》2003年第3期268-271,共4页

基　　金：国家自然科学基金资助项目 ( 9710 0 10 9)

摘　　要：应用 PCR技术 ,从小鼠基因组中克隆了 1 .6 kb乳清酸蛋白 (WAP)基因 5′上游调控序列 ,经酶切和测序证实与已知序列基本一致。为了证实此调控序列能否指导外源基因在乳腺中的表达 ,将此序列与报告基因 CAT融合 ,构建了 p WAP- CAT- poly A乳腺定位表达载体 ,脂质体包裹后 ,经乳导管将其注入到妊娠中后期家兔乳腺中进行暂时表达。分别于家兔分娩后 1～ 1 0 d采奶 ,用 EL ISA检测乳汁中 CAT含量。结果表明 ,所构建的载体在家兔乳腺中能够表达 ,表达水平在家兔分娩后 1～ 5The 5′upstream regulatory sequence of 1.6 kb(-1 573～-27)of mouse WAP gene were amplified by PCR from mouse liver DNA. The regulatory region was then inserted into the T vector. The restriction endonuclease analysis and sequence determination of the gene fragment was performed. The result showed that the regulatory element in the 5′upstream flanking region sequence was the same as the sequence published by S.M.Campbell. In order to identify the regulatory ability to direct the expression of the foreign gene in mammary gland, the mammary gland specific expression vector, pWAP CAT, was constructed using the 1.6 kb mouse WAP gene 5′flanking region fused with the report gene CAT, then, the vector was mixed with liposome and injected into the mammary gland of the later stage of gestation of pregnant rabbits through teat canal. The result of ELISA showed that the CAT gene could be expressed in rabbit mammary glands.

关 键 词：小鼠 乳清酸蛋白 上游调控序列 克隆 CAT基因 家兔 乳腺表达 

分 类 号：Q78[生物学—分子生物学]