山羊催乳素受体基因部分片段的克隆与序列分析

Cloning and Sequence Analysis of Partial Fragment of Prolactin Receptor Gene in Goats

采用PCR技术扩增出济宁青山羊催乳素受体基因长892 bp的片段,该片段含有97 bp的部分外显子8序列(外显子8全长为100 bp)、683 bp的内含子8、70 bp的外显子9及42 bp的部分内含子9。将该片段克隆到pGEM-T Easy质粒中,重组质粒用PCR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。该序列与绵羊、母牛、人、大鼠、小鼠的催乳素受体基因mRNA的对应序列的核苷酸同源性分别为99.4%、97.01%、89.22%、89.22%、88.02%,氨基酸同源性分别为100%、94.55%、81.88%、81.82%、83.64%。

The 892 bp fragment （97 bp of part of exon 8 （the full length of exon 8 was 100 bp）, 683 bp of intron 8, 70 bp of exon 9 and 42 bp of part of intron 9）of prolactin receptor （PRLR） gene was amplified successfully in Jining Grey goats by PCR and cloned into pGEM-T Easy vector. The positive clones were further identified by PCR analysis. The nucleotide sequence was detected and the peptide sequence of this fragment was deduced. This sequence shared 99.4 %, 97.01%, 89.22%, 89.22%, 88.02% nucleotide homology with the published mRNA of PRLR gene of sheep, cow, human, rat and mouse separately, and the amino acid homology was 100%, 94.55%, 81.88%, 81.82%, 83.64% separately.