双峰驼MHC-DRB基因exon2克隆序列分析及遗传多样性研究

发布时间：2018-05-05 03:34

本文选题：双峰驼 + MHC-DRB基因　；参考：《内蒙古农业大学》2015年硕士论文

【摘要】：目的：本研究通过探索内蒙古阿拉善右旗、阿拉善左旗、鄂尔多斯市、巴彦淖尔市、锡林郭勒盟地区的双峰驼群体MHC-DRB基因第二外显子序列多态性；分析了阿拉善双峰驼MHC-DRB基因与阴道蝇蛆病的相关性；研究了阿拉善双峰驼和苏尼特双峰驼两个品种遗传多样性,并判断其亲缘关系,旨在为以后双峰驼的抗病育种提供重要的信息。方法：(1)对内蒙古地区121峰双峰驼MHC-DRB基因外显子2进行克隆测序,用不同分析软件进行序列分析,确定多态位点、等位基因种类、氨基酸变异位点等。(2)采用PCR产物直接测序的方法检测了40峰患阴道蝇蛆病的阿拉善双峰驼和54峰未患阴道蝇蛆病的阿拉善双峰驼的MHC-DRB基因第二外显子序列,并计算了患病驼和未患病驼的基因频率和基因型频率。(3)采用PCR产物直接测序的方法检测了72峰阿拉善双峰驼和25峰苏尼特双峰驼MHC-DRB基因外显子2序列多态性；用分析软件计算97峰双峰驼序列的遗传多样性参数值,NJ法构建两品种系统发育树。结果：(1)运用分析软件检测了121峰双峰驼DRB基因第二外显子序列,共发现了225个突变位点、213个多态位点；36种等位基因；84个氨基酸中有62个变异位点,其中包括18个假定抗原结合位点。(2)通过对阿拉善双峰驼中患蝇蛆病驼和健康驼序列分析,发现都由两个不同的基因控制；健康驼有三种基因型,患病驼有两种基因型。(3)通过软件分析了72峰阿拉善双峰驼和25峰苏尼特双峰驼序列,发现阿拉善双峰驼有三种单倍型,苏尼特双峰驼有两种单倍型。(4)对93和159多态位点进行哈迪-温伯格平衡检验,发现P值均小于0.05；(5)双峰驼MHC-DRB基因序列进行Tajima's检验,Tajima'sD值为正值,P0.01；(6)两个双峰驼品种的MHC-DRB基因外显子2序列构建系统发育树,大多数阿拉善双峰驼聚在一支,苏尼特双峰驼聚在另一支上,也有一部分两品种双峰驼混合分布在一支上。结论：MHC-DRB基因在五个地区双峰驼群体中具有丰富的多态性；阿拉善双峰驼中患病驼和健康驼与MHC-DRB基因有一定的相关性；阿拉善双峰驼和苏尼特双峰驼MHC-DRB基因外显子2序列都存在遗传多样性,且苏尼特双峰驼的遗传多样性高于阿拉善双峰驼;两品种双峰驼亲缘关系非常近。
[Abstract]:Objective: to explore the polymorphism of the second exon of MHC-DRB gene in Bactrian camel population in Inner Mongolia Alashan right Banner, Alashan Zuo Banner, Ordos City, Bayan Nur City and Xilingol region. The relationship between MHC-DRB gene and vaginal myiasis in Alashan Bactrian Camel was analyzed, the genetic diversity of Alashan Bactrian Camel and Sunita Bactrian Camel was studied, and the genetic relationship was judged. The aim is to provide important information for disease resistance breeding of Bactrian camel in the future. Methods: 1) cloning and sequencing of exon 2 of MHC-DRB gene of 121-peak bactrian camel in Inner Mongolia. The polymorphic loci and alleles were determined by using different analysis software. Amino acid mutation site et al. (2) the MHC-DRB gene exon 2 of Alashan bactrian camel with 40 peaks of vaginal myiasis and 54 peaks without vaginal myiasis was detected by direct sequencing of PCR products. The gene frequency and genotype frequency of infected and uninfected camels were calculated. The polymorphism of exon 2 of MHC-DRB gene of Alashan Bactrian Camel and 25 Sunita Bactrian Camel was detected by direct sequencing of PCR products. The genetic diversity parameter values of 97 peak bactrian camel sequence were calculated by using the analysis software and NJ method was used to construct the phylogenetic tree of two varieties. Results the second exon sequence of DRB gene of Bactrian Camel was detected by using analysis software. A total of 225 mutation sites and 36 alleles of 213 polymorphic loci were found, and 62 mutation sites were found in 84 amino acids. This includes 18 hypothetical antigen-binding sites.) by analyzing the sequences of myiasis and healthy camels in Alashan Bactrian Camels, we found that they were both controlled by two different genes; healthy camels had three genotypes. The disease camels have two genotypes.) by software analysis of 72 peak Alashan bactrian camel and 25 peak Sunita bactrian camel sequence, we found that there are three haplotypes in Alashan bactrian camel. Sunita Bactrian Camel has two haplotypes, I. e., Hadi Weinberg equilibrium test for 93 and 159 polymorphic loci. It was found that the MHC-DRB gene sequence of Bactrian Camel (P < 0.05) was detected by Tajima's. The MHC-DRB gene exon 2 sequence of two Bactrian camel varieties was constructed by Tajima's test. Most Alashan bactrian camels were clustered in the same tree. Sunita bactrian camels gather on the other, and some of the two species are mixed in one. Conclusion there is a rich polymorphism in the population of Bactrian Camel (Bactrian Camel) in five regions, and there is a certain correlation between the MHC-DRB gene and the diseased camel and healthy camel in Alashan Bactrian Camel. There was genetic diversity in exon 2 of MHC-DRB gene in Alashan Bactrian camel and Sunita Bactrian Camel, and the genetic diversity of Sunita Bactrian Camel was higher than that of Alashan Bactrian Camel.
【学位授予单位】：内蒙古农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S824

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