猪Reg4剪切体Reg4v1和Reg4v2的cDNA克隆和组织分布

Cloning and Tissue Distribution of Reg4 Variant Reg4v1 and Reg4v2 Gene in Pig

从猪肠道组织中提取总RNA,通过RT-PCR对Reg4基因进行cDNA扩增,获得了726 bp和832 bp的两个片段。将PCR产物分别与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。同源分析结果表明,克隆的猪Reg4 Variant2基因与人、小鼠、牛的同源性分别为71.6%、63.3%、73.6%;组织分布结果显示:猪Reg4 Variant1和Reg4 Variant2基因在肠道组织各部分均有分布。克隆的猪Reg4 Variant1和Reg4 Variant2基因分别注册GenBank(Accession.FJ469910,FJ469911)。

In order to clone and analyze the gene Reg4 in the intestinal canal tissues of pig,the total RNA was extracted and mRNA sequence of Reg4 gene was amplified by RT-PCR method which used two designed primers.The PCR products were 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 nucleotide sequence of Reg4 shared 71.6%,63.3% and 73.6% homology with that of human, mouse and cattle respectively, revealing that porcine Reg4 gene had been successfully cloned in the present study. Tissue distribution analysis showed that porcine Reg4 Variantl and Reg4 Variant2 were ubiquitously expressed in various intestinal canal tissues tested. The sequence of Reg4 Variantl and Reg4 Variant2 has been submitted to GenBank （ Accession FJ469910 and FJ469911 ）.