山羊Kiss-1基因克隆、表达模式和SNPs筛选

摘要

Kiss⁃1 gene plays an important role in reproduction process of female animals. In order to investigate its function in goats, two exons’ sequences of Kiss⁃1 in Boer goat, Haimen goat and the new breed of Suhuai goat were ampli⁃fied by PCR, and these sequences were spliced using DNAStar software and blasted with its corresponding sequences of oth⁃er species. Single nucleotide polymorphisms ( SNPs) of Kiss⁃1 coding sequence in three goats breeds were screened based on DNA pool. The results showed that the full length of CDS region of Kiss⁃1 was 408 bp in three goats encoding 135 amino acids. The nucleotide sequence of Haimen goats shared 100�00% homoly with that of new breed of Suhuai goats and 98�80% similarity with that of Boer goats. Phylogenetic tree revealed that Haimen goat and the new breeding of Suhuai goat were clustered together and then clustered with Boer goat. Two SNPs were found in Kiss⁃1 coding region of three goats breeds, among which, G256A mutation was in Haimen goats and the new breed of Suhuai goats, and A175C mutation was in Boer goats and the new breed of Suhuai goats. Kiss⁃1 showed a higher expression level in lung than in muscle and other internal organs.%Kiss⁃1在雌性动物生殖过程中发挥重要的作用。为了探讨其在山羊中的功能，本研究以波尔山羊、苏淮山羊新品系和海门山羊为研究对象，通过PCR扩增、测序和序列拼接获得Kiss⁃1基因编码区序列，利用生物信息学软件分析山羊Kiss⁃1基因序列特征，RT⁃PCR试验鉴定其在山羊中的组织表达模式，采用DNA池测序筛选Kiss⁃1基因编码区SNPs位点。结果发现3种山羊Kiss⁃1基因编码区序列长度均为408 bp，编码135个氨基酸残基；海门山羊与苏淮山羊新品系Kiss⁃1核苷酸序列一致性为100％，与波尔山羊的一致性为98�80％；系统进化发育分析发现海门山羊与苏淮山羊新品系首先聚在一起，然后再与波尔山羊聚在一起；组织表达谱显示Kiss⁃1基因在肺脏中表达量相对较高，在其他内脏和肌肉组织中表达量很少；DNA池测序发现2个SNPs位点，其中A175C位点突变引起了苏氨酸到脯氨酸的突变，G256A位点突变引起了丙氨酸到苏氨酸的改变。研究结果为进一步研究山羊Kiss⁃1基因的进化、功能及其与生产性能特别是繁殖性能的关系提供一定的理论依据。