猪Reg3基因的克隆与序列分析

Molecular Cloning and Sequence Analysis of Porcine Gene Reg3

在猪的肠道组织中克隆Reg3基因,提取总RNA,利用设计的引物进行RT-PCR,PCR产物与pMD-19T载体连接后转化E.coliJM109,检测阳性克隆、测序并进行序列分析。结果表明:克隆的猪Reg3基因与人、绵羊、马的同源性分别为83.7%,82.3%,82.2%;克隆了猪Reg3全长基因并注册GenBank注册号为Accession.FJ531494。

In order to clone and analyze the gene Reg3 in intestinal canal tissues of pig,the total RNA was extracted and mRNA sequence of Reg3 gene was amplified by RT-PCR method which used two designed primers.The PCR productswere ligated into the pMD-19T vector,and then transformed into competent cells of E.coli JM109.The sequence was analyzed to identify the recombinant plasmid.The identification analysis showed that the Reg3 nucleotide sequence shared 83.7%,82.3% and 82.2% homology with that of human,sheep and horse respectively,revealing porcine Reg3 gene had been successfully cloned in the present study.The sequence has been submitted to GenBank（Accession FJ531494）.