| 摘要: |
| 利用PCR方法克隆了牛β-乳球蛋白基因5′调控成分（1 449 bp），其中包括5′侧翼序列（645 bp），第一外显子及第一内含子（804 bp）。PCR产物回收纯化后，克隆在pMD 18-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98.55%，99.79%，90.70%和90.09%。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，可用于构建乳腺特异性表达载体。 |
| 关键词: β-乳球蛋白基因 牛 PCR 序列分析 |
| DOI： |
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| 基金项目:国家“863”高技术资助项目(2001AA213081) |
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| The isolation,cloning and sequence analysis of bovine β-lactoglobulin gene 5′regulatory elements |
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| Abstract: |
| 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. |
| Key words: β-lactoglobulin gene bovine PCR sequence analysis |
