山羊肌肉差异表达miRNA基因多态性分析
毕业论文关键词：DNA提取，PCR，SSCP，测序，多态性
Ploymorphism analysis of miRNA gene’s differential expression in goat muscle
Abstract: We had three varieties of goats to conduct the study . They are Xuhuai goat (XH , n=45) , Boer goat (BE , n=45) and Haimen goat (HM , n=45) . The technique of PCR-SSCP was taken to do the research about genetic variation on 19 miRNAs related to the development of muscle . Finally , we found that there are only 5 of them indicated ploymorphism , and they are miR-424-5p、miR-411-5p、NovelmiR-47、miR-541-3p and miRNA-767 . In this study , polymorphicsm information content turned to be accordant , just various at various loci . Apart from miRNA-411c-3p low polymorphism, the other 4 loci are medium polymorphism . At the same time, in the five polymorphic loci , most loci of the three varieties were in Hardy-Weinberg equilibrium, only Novell miR-47 and miR-767 in Haimen goat and Boer goat and miR-541-3p in Boer goat in Hardy Weinberg non-equilibrium state. In this study, miR-411-5p and miR-767 was detected with two type , AA type and AB type , but no BB type.
Key words: DNA extraction , PCR , SSCP , oder-checking , polymorphism
目录
1 引言 1
2 实验材料 2
2.1 实验动物及血样来源 2
2.2 实验试剂 2
2.3 仪器设备 2
3 试验方法 2
3.1 羊全血基因组DNA的提取 2
3.2 PCR反应所需引物设计 2
3.3 PCR-SSCP DNA标记分析 3
3.4 琼脂糖凝胶电泳检测PCR产物 4
3.5 聚丙烯酰胺凝胶电泳和检测 4
3.6 测序 4
3.7 统计分析方法 4
4 结果与分析 4
4.1 基因组DNA样品检测 4
4.2 候选miRNA遗传变异检测 4
4.2.1 NovelmiR-47基因遗传变异检测 5
4.2.2 miR-541-3p基因遗传变异检测 6
4.3 候选基因多态基因座的群体遗传学分析 7
4.3.1 NovelmiR-47前体基因群体遗传结构分析 7
4.3.2 miR-541-3p前体基因群体遗传结构分析 8
5 讨论 9
致谢 10
参考文献 10
1 引言
基因多态性在个体的繁殖过程中演绎了一个十分重要的角色，其导致了转录和翻译的多样性，这一特征十分有利于同时保证遗传的准确性与宽容度，这对于生物界的稳定和多样化起到了至关重要的作用。miRNA是一类非编码单链小RNA，其长度大约为21~23nt，它们在进化上相对保守。miRNA通过靶向mRNA来使其降解，或抑制其翻译，进而阻止靶蛋白形成。miRNA在各项生物学过程中发挥着重要的调节作用，并且在许多疾病中也都发现了miRNA的异常表达，由此也说明，基因的多态性与各种疾病有着密不可分的联系。
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