牛β-乳球蛋白基因5′调控成分的分离、克隆及序列分析  

The isolation,cloning and sequence analysis of bovine β-lactoglobulin gene 5′regulatory elements

作　　者：石玉强[1] 张涌[1] 郑月茂[1] 刘建忠[1] 张志平[1] 

机构地区：[1]西北农林科技大学生物工程研究所,陕西杨陵712100

出　　处：《西北农林科技大学学报（自然科学版）》2003年第3期1-4,共4页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"8 63"高技术资助项目 ( 2 0 0 1AA2 13 0 81)

摘　　要：利用 PCR方法克隆了牛 β-乳球蛋白基因 5′调控成分 (1 4 4 9bp) ,其中包括 5′侧翼序列 (6 4 5 bp) ,第一外显子及第一内含子 (80 4 bp)。PCR产物回收纯化后 ,克隆在 p MD1 8- T Vector的 T位点。序列分析表明 ,该序列与牛 β-乳球蛋白 A型基因、牛 β-乳球蛋白 B型基因、山羊和绵羊 β-乳球蛋白基因的同源性分别为 98.5 5 % ,99.79% ,90 .70 %和 90 .0 9%。计算机分析表明 ,此调控成分含有诸多调节因子结合位点或反应元件 ,其中包括视黄酸反应元件 (RARE)、佛波酯反应元件 (TRE)、乳腺特异性因子 (MSBF)和核因子 1 (NF1 )结合位点等 ,提示此调控成分可调控外源基因在乳腺细胞中高效表达 。For the purpose of constructing mammary gland specific expressional vector,the bovine β lactoglobulin gene 5′ flanking fragment was cloned by PCR amplification.It consisted in part of the gene upstream region about 645 bp,the first exon and the first intron about 804 bp.After PCR product was recovered and purified,the aim fragment was cloned at T site of pMD 18 T Vector.The fragment was sequenced,and it was compared with bovine BLG gene variant A,bovine BLG gene variant B,goat and sheep BLG gene.The results indicated the homology was 98.55%,99.79%,90.70% and 90.09% respectively.The nucleotide acid sequence was analyzed by computer,and found many factor or protein binding sites and response elements such as mammary specific binding factor(MSBF),nuclear factor 1 (NF 1),retinoic acid response element (RARE) and TPA response element (TRE).It suggested the cloned regulatory element should direct extra genes to express specifically in mammary gland epithelia cells,and it could be used as constructing mammary gland specific expressional vector.

关 键 词：牛 Β-乳球蛋白基因 PCR 序列分析 基因调控 乳腺生物反应器 生长激素基因 基因分离 基因克隆 

分 类 号：Q789[生物学—分子生物学] TQ464.7[化学工程—制药化工]