从江香猪抑制素α基因克隆及其卵巢表达研究

Cloning of Inhibin-αGene and its mRNA Expression Pattern in the Ovary of Congjiang Xiang Pig

为了探索抑制素α基因(inhibin-α,INHA)与从江香猪繁殖性状之间的相关性,试验采用特异性聚合酶链式反应(PCR)技术克隆从江香猪INHA基因,测定其核苷酸序列,通过等位基因特异性PCR(allele-specific PCR,AS-PCR)方法检测从江香猪低产群与高产群之间INHA基因的多态性变化,以实时荧光定量PCR技术检测高产、低产从江香猪卵巢组织中INHA基因的表达量。结果表明,从从江香猪基因组中成功克隆了INHA基因,编码区完整,全长1 095bp,编码364个氨基酸;经比对发现,INHA基因外显子2序列中存在2个候选SNPs位点(G359A和A373G)。经大样本检测,从江香猪高产群与低产群之间2个候选SNPs位点的基因频率没有明显差异;相比之下,高产从江香猪卵巢中INHA基因的表达量较高。研究结果提示,从江香猪INHA基因结构保守,可能主要通过基因的表达量变化调节从江香猪卵巢的生长和卵泡的发育。

To reveal the relationship between inhibin-α（INHA）gene and the reproductive traits of Congjiang Xiang pig,INHAgene was cloned and sequenced taking the genomic DNA of Congjiang Xiang pigs as templates by polymerase chain reaction（PCR）method.The polymorphisms of INHAgene were tested in Congjiang Xiang pig populations with high-litter size and low-litter size using allele-specific PCR（AS-PCR）method.The expression profile of INHAgene in ovaries was detected from Congjiang Xiang pigs with high-litter yiled or low-litter yiled by Real-time PCR method.The complete coding region of INHAgene was 1 095 bp in length,which coded for 364 amino acid residues.Compared with the known sequence,two candidate sites,G359 Aand A373G,were found out from exon 2region of INHAgene in Congjiang Xiang pig.After investigation for the two sites in a large population,the frequency of alleles between two populations was not significant and without obvious relativity with the litter yiled of Congjiang Xiang pig.However,the INHA mRNA level in the ovary of Congjiang Xiang pig with high-litter yiled was higher than that with low-litter yiled.It suggested that INHAgene was much conserved,INHAgene expressionlevel might be concerned for the regulation of ovary growth and follicle development in Congjiang Xiang pig breed.