北京荷斯坦母牛BoLA-DRB3基因外显子2的克隆与序列分析

Cloning and Sequence Analysis of Exon 2 of BoLA-DRB3 Gene in Beijing Holstein Cows

采用PCR技术扩增出北京荷斯坦母牛BoLA-DRB3基因的外显子2的267 bp大小的片段,将该片段克隆到pGEM-T Easy质粒中,重组质粒用PCR和酶切分析进行阳性克隆鉴定,然后测定核苷酸序列,并推导其氨基酸序列.北京荷斯坦牛、蒙古牛、人类、瑞典红白花牛、蒙古绵羊、西班牙山羊之间DRB3基因外显子2的核苷酸序列同源性为80.9%～95.5%,氨基酸序列同源性为75.3%～91.0%.

The 267 bp fragment of exon 2 of BoLA-DRB3 gene was amplified successfully by PCR and cloned into pGEM-T Easy vector. The positive clone was further identified by PCR and restriction enzyme analysis. The nucleotide sequence was detected and the peptide sequence of this fragment was deduced. Homologies of nucleotide sequence and amino acid sequence of exon 2 of DRB3 gene among Beijing Holstein cattle, Mongolian cattle, human, Swedish Red and White cattle, Mongolian sheep and Spanish goat were from 80.9% to 95.5 % and from 75.3% to 91.0%, respectively.