牛酪蛋白基因的长程限制酶切分析

Ferretti L, Leone P, Sgaramella V

IDVGA, CNR, Milan, Italy.

Nucleic Acids Res. 1990 Dec 11;18(23):6829-33. doi: 10.1093/nar/18.23.6829.

Pulsed field gel electrophoresis (PFGE) was used to analyse the organization of the bovine alpha s1, alpha s2, beta and kappa casein genes. High molecular weight DNA was prepared from fibroblasts and lymphocytes embedded in agarose and was digested with the restriction endonucleases Clal, Sall, Smal, Xhol. The digestion products were separated by PFGE, transfered to nitrocellulose filters and hybridized to probes corresponding to the cDNAs of the four bovine casein genes. The casein genes were demonstrated to be physically linked within a region of 300 kb, represented by two adjacent Xhol fragments in fibroblasts and by a single fragment in lymphocytes. A restriction map of the casein locus was derived and the order of the genes was shown to be kappa, alpha s2, beta, alpha s1.

脉冲场凝胶电泳（PFGE）用于分析牛αs1、αs2、β和κ酪蛋白基因的组织。从包埋在琼脂糖中的成纤维细胞和淋巴细胞中制备高分子量DNA，并用限制性内切酶ClaI、SalI、SmaI、XhoI进行消化。消化产物通过PFGE分离，转移到硝酸纤维素滤膜上，并与对应于四个牛酪蛋白基因cDNA的探针杂交。结果表明，酪蛋白基因在300kb的区域内物理连锁，在成纤维细胞中由两个相邻的XhoI片段代表，在淋巴细胞中由一个片段代表。推导了酪蛋白基因座的限制性图谱，基因顺序显示为κ、αs2、β、αs1。