鹅催乳素受体基因3′-末端序列克隆

Cloning of 3′-end of Prolactin Receptor Gene in Goose

通过比对鸡、火鸡、鸽、猪、大鼠5种动物及人催乳素受体(PRLR),并根据它们的基因cDNA保守区设计兼并引物,先扩增了鹅催乳素受体基因(gPRLR)cDNA一段606 bp保守区序列,再以此设计基因特异性引物,利用3′-RACE技术克隆了gPRLRcDNA3′-末端序列。序列分析表明,获得的gPRLRcDNA 3′-末端长1 876 bp,含编码区后1 656 bp的编码核苷酸、终止密码子TAA和220 bp的3′-UTR。参照鸡催乳素受体基因(cPRLR),此gPRLRcDNA3′-末端序列可分成6个外显子,其长度分别为148 bp、170 bp、145 bp、100 bp、70 bp和1 243 bp,与cPRLRcDNA最后6个外显子高度同源,终止密码子位于最后1个外显子的1 021 bp处。编码区后1 653 bp编码的551个氨基酸gPRLR C-末端与鸡PRLR同源部分的同源性达到87.7%。

To characterize the prolactin receptor gene family of goose,a eDNA fragment of goose prolactin receptor gene （gPRLR） with a length of 606 bp was first isolated by RT-PCR using a pair of degenerate primers which were designed based on conservative regions among six species incluing of human, chicken, turkey, pigeon, pig and rat. The 3＇-end of gPRLR eDNA was identified by 3 ＇-RACE strategy using gene-specific primers complementary to the isolated eDNA fragment. Sequence analysis of the 1 876 bp RACE product showed a 1 656 bp 3＇coding region, a stop codon （TAA） and a 220 bp 3＇-UTR. Six predicted exons with sizes of 148 bp, 170 bp, 145 bp, 100 bp, 70 bp and 1 243 bp, respectively, which were found in the sequences were highly homologous to the last exons of cPRLR eDNA. The C-terminus of gPRLR with 551 amino acids long encoded by the last 1 653 bp of the region exhibited 87.7% similarity to the corresponding region of chicken prolactin receptor. Stop codon was located at 1 021 bp of last exon.