马可·波罗盘羊和5种家养绵羊MHC-DRB3基因第2外显子遗传多态性分析

doi:10.16431/j.cnki.1671-7236.2022.03.018

摘要/Abstract

摘要： 【目的】分析马可·波罗盘羊MHC-DRB3基因第2外显子多态性，确定其等位基因数及核苷酸、氨基酸多态位点，以及与家养绵羊的遗传进化关系。【方法】采用PCR-SSCP方法对39只马可·波罗盘羊、159只塔什库尔干羊、72只藏绵羊、76只宁夏滩羊、36只柯尔克孜羊和26只多浪羊MHC-DRB3基因第2外显子的等位基因进行检测，对不同绵羊品种产生的差异等位基因进行克隆及测序，并对等位基因单倍型序列进行分析；利用生物信息学在线软件分析马可·波罗盘羊MHC-DRB3基因第2外显子编码蛋白的理化性质、结构和功能。【结果】试验共获得66个MHC-DRB3等位基因，马可·波罗盘羊、塔什库尔干羊、藏绵羊、宁夏滩羊、柯尔克孜羊和多浪羊的等位基因数分别为23、14、12、12、4和1个。马可·波罗盘羊、塔什库尔干羊、藏绵羊、宁夏滩羊和柯尔克孜羊MHC-DRB3基因第2外显子等位基因序列中的核苷酸多态位点分别为74、76、45、49和25个，表明相较于藏绵羊、宁夏滩羊和柯尔克孜羊，马可·波罗盘羊和塔什库尔干羊的多态性更高。系统发育分析表明，马可·波罗盘羊和塔什库尔干羊MHC-DRB3基因均呈两支分化；马可·波罗盘羊、塔什库尔干羊和藏绵羊MHC-DRB3基因单倍型的相似性较高。马可·波罗盘羊MHC-DRB3基因第2外显子编码产物为不稳定亲水性蛋白，二级结构主要以无规则卷曲和α-螺旋为主，其生物学作用主要在细胞质中体现。【结论】本试验确定了马可·波罗盘羊和5种家养绵羊MHC-DRB3基因第2外显子的等位基因数及核苷酸、氨基酸多态位点，为进一步探究绵羊MHC-DRB3基因的调控机理及进化意义提供材料。

关键词: 马可·波罗盘羊; 家养绵羊; MHC-DRB3基因; PCR-SSCP; 遗传多态性

Abstract: 【Objective】 This study was aimed to analyze the polymorphism of MHC-DRB3 gene in Ovis ammon polii, determine its allele number, nucleotide and amino acid polymorphic loci, as well as genetic relationship and evolutionary relationship with domestic sheep.【Method】 PCR-SSCP method was used to detect the alleles of MHC-DRB3 gene exon 2 in 39 Ovis ammon polii, 159 Taxkorgan sheep, 72 Tibetan sheep, 76 Ningxia Tan sheep, 36 Kirgiz sheep and 26 Duolang sheep.The differential alleles of different sheep breeds were cloned and sequenced, and the haplotype sequences of alleles were analyzed.The physical and chemical properties, structure and function of MHC-DRB3 gene exon 2 coded protein in Ovis ammon polii were analyzed by bioinformatics online softwares.【Result】 This study obtained 66 MHC-DRB3 gene alleles, including 23 alleles for Ovis ammon polii, 14 alleles for Taxkorgan sheep, 12 alleles for Tibetan sheep, 12 alleles for Ningxia Tan sheep, 4 alleles for Kirgiz sheep, and 1 allele for Duolang sheep.The number of nucleotide polymorphic loci of allele sequence of MHC-DRB3 gene exon 2 in Ovis ammon polii, Taxkorgan sheep, Tibetan sheep, Ningxia Tan sheep and Kirgiz sheep were 74, 76, 45, 49 and 25, respectively.It showed that Ovis ammon polii and Taxkorgan sheep had higher polymorphisms than Tibetan sheep, Ningxia Tan sheep and Kirgiz sheep.The phylogenetic analysis results showed that the MHC-DRB3 gene of Ovis ammon polii and Taxkorgan sheep both diverged into two branches.The MHC-DRB3 gene haplotypes of Ovis ammon polii, Taxkorgan sheep and Tibetan sheep had high similarity.The product encoded by MHC-DRB3 gene exon 2 of Ovis ammon polii was an unstable hydrophilic protein.The secondary structure mainly composed of random coil and alpha helix, and its biological effects were mainly reflected in cytoplasm.【Conclusion】 In this study, the number of alleles of MHC-DRB3 gene exon 2 and the polymorphic loci of nucleotide and amino acid of MHC-DRB3 gene in Ovis ammon polii and five species of domestic sheep were determined, it provided materials for further exploring the regulatory mechanism and evolutionary significance of MHC-DRB3 gene in sheep.

Key words: Ovis ammon polii; domestic sheep; MHC-DRB3 gene; PCR-SSCP; genetic polymorphism

中图分类号:

S813.3

张永浩, 肖逸, 郭丽君, 阿布都热合曼·吐尔逊, 王玉涛. 马可·波罗盘羊和5种家养绵羊MHC-DRB3基因第2外显子遗传多态性分析[J]. 中国畜牧兽医, 2022, 49(3): 962-972.

ZHANG Yonghao, XIAO Yi, GUO Lijun, Tuerxun·Abudoureheman, WANG Yutao. Genetic Polymorphism Analysis of MHC-DRB3 Gene Exon 2 in Ovis ammon polii and Five Domestic Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 962-972.

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