Ahn J Y, Aoki N, Adachi T, Mizuno Y, Nakamura R, Matsuda T
Department of Applied Biological Sciences, School of Agricultural Sciences, Nagoya University, Japan.
Biosci Biotechnol Biochem. 1995 Jan;59(1):59-64. doi: 10.1271/bbb.59.59.
Bovine mammary epithelial cells (BMEC) were isolated as acinous fragments from a mammary gland of a lactating cow. They grew well on plastic substratum, showed the characteristic cobblestone morphology of epithelial cells, and secreted alpha s1-, beta-, and kappa-caseins even when grown on plastic substratum. A plasmid containing the bacterial chloramphenicol acetyl transferase (CAT) gene was transfected to the isolated BMEC by calcium phosphate precipitation and electroporation methods. The transfection efficiency of BMEC by the calcium phosphate method was greatly improved by post-transfection osmotic shock with glycerol or polyethylene glycol. An about 700 bp DNA fragment containing 5'-flanking sequence of bovine alpha s1-casein gene showed promoter activity in the transfected BMEC. The primary culture of BMEC might be useful for studies on regulation of bovine milk-protein gene expression.
牛乳腺上皮细胞（BMEC）从泌乳期奶牛的乳腺中分离为腺泡片段。它们在塑料基质上生长良好，呈现上皮细胞特有的鹅卵石形态，即使在塑料基质上生长时也能分泌αs1-、β-和κ-酪蛋白。通过磷酸钙沉淀法和电穿孔法将含有细菌氯霉素乙酰转移酶（CAT）基因的质粒转染到分离的BMEC中。用甘油或聚乙二醇进行转染后渗透休克可大大提高磷酸钙法对BMEC的转染效率。一个包含牛αs1-酪蛋白基因5'-侧翼序列的约700 bp DNA片段在转染的BMEC中显示出启动子活性。BMEC的原代培养可能有助于研究牛乳蛋白基因表达的调控。
