绵羊鸟苷素基因的克隆与序列分析

基于电子延伸序列,本试验克隆并分析了绵羊鸟苷素基因.从绵羊十二指肠黏膜组织中提取总RNA,利用设计的引物进行RT-PCR扩增,PCR产物与pMD19-T载体连接后转化到E.coliJM109感受态细胞,检测阳性克隆并测序.结果表明:克隆的绵羊鸟苷素基因长341 bp,编码109个氨基酸,与人、豚鼠、小鼠和牛的同源性分别为77%7、5%、47%和94%,推测的氨基酸序列信号肽为1~16 aa,整个氨基酸构成一个模域,编码鸟苷素前体蛋白.

Molecular cloning and sequence analysis of ovine guanylin

The coding sequence of ovine guanylin was cloned and analyzed based on the in silico sequence information.Total RNA was extracted from mucosal tissue of ovine duodenum and mRNA sequence was amplified by RT-PCR with designed primers.The PCR products were ligated into the pMD19-T vector,and then transformed into competent E.coli JM109.The sequence was analyzed to identify the recombinant plasmid.Sequence analysis indicated that the guanylin gene was 341 bp in length and the deduced amino acids was 109.Homology analysis showed that the sequence shared 77 %,75 %,74 % and 94 % homology with those of human,guinea pig,mouse and cattle.The signal peptide of deduced amino acid sequence is 1～16 aa and the omain is formed by the amino acid sequence coding precursor protein of guanylin.