【作者】 邢凤；

【导师】 王建民；

【作者基本信息】 山东农业大学 ， 动物遗传育种与繁殖， 2008， 硕士

【摘要】 主要组织相容性复合物(MHC)是一群紧密连锁高度多态的基因,编码主要组织相容性抗原,在脊椎动物机体的免疫系统中具有重要作用,因此MHC常作为家畜抗病育种的候选基因。本研究采用PCR-SSCP、PCR-RFLP技术对莱芜黑山羊、波尔山羊、鲁波杂交山羊3个山羊种群共175只个体的DQA1、DQA2、DQB1基因外显子2进行了遗传多态性分析,并对DQB1、DQA2基因多态性与线虫抗性、免疫性状的相关性进行了研究,其结果如下:1.采用PCR-SSCP技术对DQA1基因外显子2多态性进行了分析,共检测到8种基因型,并对其纯合子进行了测序,检测到23处碱基突变。2.对人类、山羊、绵羊、牛DQA1基因外显子2核苷酸序列进行同源性比较,结果发现同源性很高。3.采用PCR-RFLP技术对莱芜黑山羊、鲁波杂交山羊和波尔山羊DQA2、DQB1基因外显子2进行多态性分析,共检测到4种DQA2基因外显子2基因型,7种DQB1基因外显子2基因型。通过酶切图谱和测序分析表明,DQA2基因外显子2的第77、79、80和169位的碱基表现出多态性,DQB1基因外显子2的第24、151位的碱基表现出多态性。4.应用SAS 8.0软件GLM程序对DQA2、DQB1基因不同基因型与线虫抗性进行了相关分析,在DQA2基因中,基因型与线虫EPG有一定的相关性,但没有达到显著水平(P>0.05)。在DQB1基因中,BC、DD基因型的线虫EPG显著低于BB基因型(P<0.05),在种群内的分析中发现,鲁波杂交山羊中,BC基因型的线虫EPG显著低于AC基因型(P<0.05)。其它基因型与线虫EPG也有一定的相关性,但没有达到显著水平(P>0.05)。5.应用SAS 8.0软件GLM程序对DQA2、DQB1基因不同基因型与免疫性状进行了相关分析,在DQA2基因中,莱芜黑山羊中AB基因型的RBC低于BB、BC基因型(P>0.05),而MCH高于BB、BC基因型,但没有达到显著水平(P>0.05);BC基因型的W-LCR高于AB、BB基因型(P>0.05),而W-SCC低于AB、BB基因型,但没有达到显著水平(P>0.05)。鲁波杂交山羊中AB基因型的RBC显著高于BB基因型(P<0.05),高于BC基因型(P>0.05);AB基因型的WBC显著高于BC基因型(P<0.05),高于BB基因型(P>0.05);AB基因型的HCT显著高于BB、BC基因型(P<0.05);BC基因型的W-LCR显著高于BB基因型(P<0.05),高于AB基因型(P>0.05)。在DQB1基因中,莱芜黑山羊中BC基因型的W-SCR显著高于AC、CC基因型(P<0.05),W-LCR显著低于CC基因型(P<0.05),W-LCC低于AC、CC基因型,但差异不显著(P>0.05);波尔山羊中BC基因型的W-LCC低于AA、AB基因型,但差异不显著(P>0.05);鲁波杂交山羊中BC、AC基因型的W-LCC显著低于AA基因型(P<0.05)。更多还原

【Abstract】 MHC is a chromosomal region consisting of a group of closely linked loci which are highly polymorphic and play a central role in the immune system.So it is a candidate gene in disease resistance breeding. PCR-SSCP/PCR-RFLP were used to analyze the polymorphisms of exon 2 of DQA1,DQA2,DQB1 gene in Laiwu black goats, Lubo goats and Boer goats.The effects ofgenotypes of exon 2 of DQB1,DQA2 gene on nematode resistance ,immune traits were estimated.The results were as followed:1. Polymorphisms of exon 2 of DQA1 was analyzed by PCR-SSCP , Eight genotypes were detected.and Its homozygous were sequenced, 23 polymorphic sites were detected .2. The exon 2 of DQA1gene were amplified by PCR respectively. DNA sequences are compared. Comparative analysis showed that the homologies of DQA1 gene among Human, Bovine, sheep, goats, Laiwu black goats are very high.3. PCR-RFLP was used to analyze the polymorphisms of exon 2 of DQA2 ,DQB1 gene in Laiwu black goats,Lubo goats and Boer goats.it was showed four DQA2 gene exon 2 genotypes and seven DQB1 gene exon 2 genotypes by analyzing restriction map and sequencing. Polymorphic sites were detected at base position 77, 79,80,169 of exon 2 of DQA2 gene. 24,151 of exon 2 of DQB1 gene.4. The association of polymorphisms of exon 2 of DQA2, DQB1 gene with nematode resistance in three goat breeds was analyzed by SAS 8.0 software GLM program, In DQB1 gene, the nematode eggs count of genotype BC, DD was significantly lower than that with genotype BB (P <0.05). by analysising in the breeds , we found that in Lubo goats, the nematode eggs count with genotype BC was significantly lower than that with genotype AC (P <0.05). other genotypes and nematode eggs count had some relevance, but did not reach significant levels (P> 0.05).5. The association of polymorphisms of exon 2 of DQA2 , DQB1 gene with immune traits in three goats was analyzed by SAS 8.0 software GLM program. The results suggested that for DQA2 gene, RBC in Laiwu black goats with genotype AB was lower than that with genotype BB,BC(P>0.05), MCH in Laiwu black goats with genotype AB was higher than that with genotype BB, BC(P>0.05);W-LCR in Laiwu black goats with genotype BC was higher than that with genotype AB,BB(P>0.05);W-SCC in Laiwu black goats with genotype BC was lower than that with genotype AB, BB(P>0.05). RBC in Lubo goats with genotype AB was significantly higher than that with genotype BB (P<0.05), RBC in Lubo goats with genotype AB was higher than that with genotype BC(P>0.05). WBC in Lubo goats with genotype AB was significantly higher than that with genotype AC (P<0.05). WBC in Lubo goats with genotype AB was higher than that with genotype BB(P>0.05).HCT in Lubo goats with genotype AB was significantly higher than that with genotype BB ,BC (P<0.05).W-LCR in Lubo goats with genotype BC was significantly higher than that with genotype BB (P<0.05). W-LCR in Lubo goats with genotype BC was higher than that with genotype AC(P>0.05). for DQB1 gene,W-SCR in Laiwu black goats with genotype BC was significantly higher than that with genotype AC and CC (P<0.05), W-LCR in Laiwu black goats with genotype BC was significantly lower than that with genotype CC (P<0.05); W-LCC in Laiwu black goats with genotype BC was lower than that with genotype AC, CC (P>0.05). W-LCC in Boer goats with genotype BC was lower than that with genotype AA , AB (P>0.05) ; W-LCC in Lubo goats with genotype BC and AC was significantly lower than that with genotype AA (P<0.05).These results may be applied to marker assisted selection in disease resistance breeding of goats.更多还原