摘要
在猪的不同组织中克隆KISS-1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.coliDH5α,检测阳性克隆并测序;结果:克隆的猪KISS-1基因与牛、人、大鼠、小鼠同源性分别为81.0%、76.8%、71.4%、72.4%;克隆了猪KISS-1基因全长并注册GenBank(Accession.EU934235),猪KISS-1基因在多种组织中表达。
In order to clone and analyze the gene KISS - 1 in various tissues of pig, the total RNA was extracted and mRNA sequence of KISS - 1 gene was amplified by RT - PCR method which used two designed primers. The PCR products were ligated in-to the pMD-19Tveetor, and then transformed into competent cells of E. coli DH5α. The sequence was analyzed to identify the re- combinant plasmid. The identification analysis showed that the KISS - 1 nucleotide sequence shared 81.0%, 76.8%, 71.4% and 72.4% homology with that of Bos taurus, human , rat and mouse respectively, revealing porcine KISS-1 gene had been successfully cloned in the present study. The sequence has been submitted to GenBank （ Accession EU934235 ）. The expression of porcine KISS - 1 gene in various tissues was found.
出处
《江西农业学报》
CAS
2008年第11期5-7,共3页
Acta Agriculturae Jiangxi
基金
河南省重点科技攻关项目(0523010500)
