Sun Dong-Xiao, Zhang Yuan, Li Ning
College of Animal Science and Technology, China Agricultural University, Beijing 100094, China.
Yi Chuan Xue Bao. 2003 Aug;30(8):761-5.
MHC(major histocompatibility complex) is a chromosomal region consisting of a group of closely linked loci which are highly polymorphic, which plays a central role in the immune system of animals and is almost found in all vertebrates. Its expressional product is MHC antigen of which main function is antigen presentation. In this study, the exon 2 of MHC-DRB3 gene was amplified by PCR and a uniform fragment of 285 bp was obtained in Mongolian Sheep and Kazakh Sheep. The 285 bp PCR product was digested with restriction endomuclease HaeIII and genetic polymorphism was investigated by PCR-RFLP. Seven alleles were found: A(168 bp/117 bp), B(154 bp/117 bp/14 bp), C(154 bp/65 bp/52 bp/14 bp), D(168 bp/65 bp/52 bp), E(154 bp/131 bp), F(154 bp/66 bp/65 bp), H(220 bp/65 bp). There are seventeen genotypes in the tested populations, BB(154 bp/117 bp/14 bp), CC(154 bp/65 bp/52 bp/14 bp), EE(154 bp/131 bp), HH(220 bp/65 bp), FF(154 bp/66 bp/65 bp), AB(168 bp/154 bp/117 bp/14 bp), AC(168 bp/154 bp/117 bp/65 bp/52 bp/14 bp), AE(168 bp/154 bp/131 bp/117 bp), AH(220 bp/168 bp/117 bp/65 bp), BC(154 bp/117 bp/65 bp/52 bp/14 bp), CE(154 bp/131 bp/65 bp/52 bp/14 bp), CD(168 bp/154 bp/65 bp/52 bp/14 bp), CF(154 bp/66 bp/65 bp/52 bp/14 bp), CH(220 bp/154 bp/65 bp/52 bp/14 bp), DE(168 bp/154 bp/131 bp/65 bp/52 bp), EH(220 bp/154 bp/131 bp/65 bp), FH(220 bp/154 bp/66 bp/65 bp). By analyzing restriction map, polymorphic sites were detected at base position 154, 168 and 220. Statistical results indicated that there are significant differences of some genotypic frequencies and allelic frequencies between Mongolian Sheep and Kazakh Sheep(P < 0.10, P < 0.05 or P < 0.01). CC, FF, BC and B, C, F are favorable genotypes and favorable alleles respectively in Mongolian Sheep and Kazakh Sheep, In addition, HaeIII CE and HaeIII E is favorable genotype and favorable allele respectively in Kazakh Sheep. The results of chi 2 test showed that genotypes of MHC-DRB, gene in Mongolian Sheep and Kazakh Sheep did not fit with Hardy-Weinberg equilibrium(P < 0.01).
主要组织相容性复合体(MHC)是一个染色体区域,由一组紧密连锁且高度多态的基因座组成,在动物免疫系统中起核心作用,几乎存在于所有脊椎动物中。其表达产物是MHC抗原,主要功能是呈递抗原。本研究通过PCR扩增了蒙古羊和哈萨克羊MHC-DRB3基因的外显子2,获得了一条285bp的一致片段。用限制性内切酶HaeIII对285bp的PCR产物进行酶切,采用PCR-RFLP技术研究其遗传多态性。共发现7个等位基因:A(168bp/117bp)、B(154bp/117bp/14bp)、C(154bp/65bp/52bp/14bp)、D(168bp/65bp/52bp)、E(154bp/131bp)、F(154bp/66bp/65bp)、H(220bp/65bp)。被测群体中有17种基因型,BB(154bp/117bp/14bp)、CC(154bp/65bp/52bp/14bp)、EE(154bp/131bp)、HH(220bp/65bp)、FF(154bp/66bp/65bp)、AB(168bp/154bp/117bp/14bp)、AC(168bp/154bp/117bp/65bp/52bp/14bp)、AE(168bp/154bp/131bp/117bp)、AH(220bp/168bp/117bp/65bp)、BC(154bp/117bp/65bp/52bp/14bp)、CE(154bp/131bp/65bp/52bp/14bp)、CD(168bp/154bp/65bp/52bp/14bp)、CF(154bp/66bp/65bp/52bp/14bp)、CH(220bp/154bp/65bp/52bp/14bp)、DE(168bp/154bp/131bp/65bp/52bp)、EH(220bp/154bp/131bp/65bp)、FH(220bp/154bp/66bp/65bp)。通过分析酶切图谱,在第154、168和220位碱基处检测到多态性位点。统计结果表明,蒙古羊和哈萨克羊的一些基因型频率和等位基因频率存在显著差异(P<0.10、P<0.05或P<0.01)。CC、FF、BC以及B、C、F分别是蒙古羊和哈萨克羊的优势基因型和优势等位基因。此外,HaeIII CE和HaeIII E分别是哈萨克羊的优势基因型和优势等位基因。卡方检验结果表明,蒙古羊和哈萨克羊MHC-DRB基因的基因型不符合哈迪-温伯格平衡(P<0.01)。