中国少数民族基因组DNA样本库的建立和CYP2D6在维吾尔族人群的多态性分布

发布时间：2018-11-14 21:14

【摘要】：[目的与意义] 随着中国社会政治和经济的发展,各民族融合日益加剧,纯系少数民族遗传资源日渐珍贵。特别是少数民族遗传资源外流初露端倪,亟需加大少数民族遗传资源的保护工作。故此,我们建立包含中国56个民族遗传信息的大样本的基因组DNA样本库,以充分保存和保护现有的遗传资源。这些遗传资源可以为人类提供种族的起源,进化及迁徙等信息,同时对于开展基于遗传因素的药效学和药物代谢动力学,甚至一些遗传疾病发生机制的研究有极重要的意义。近年来,遗传药理学的研究成为了医学领域研究的热点,尤其是遗传因素与药物不良反应的发生之间的研究,世界各国都给予了不同程度的关注和重视。药物代谢酶的多态性的存在,是导致药物药效和不良反应个体差异的主要原因。我国各少数民族人群均有自己独特的遗传特质,势必会导致同一药物在不同民族人群产生不同的疗效,甚至发生不良反应。因此,了解各民族药物代谢水平的差异是优化治疗剂量,减少药物不良反应以及推广少数民族传统药物的关键因素之一。有分析显示,由P450的多态性造成的不良反应占药物不良反应总数的86%。细胞色素P4502D6 (CYP2D6)是一种重要的最具多态性的药物代谢酶之一,代谢了临床上约20-30%的药物,由CYP2D6的多态性导致的药物不良反应和药物代谢差异已有较多的报道。本研究旨在建立中国少数民族基因组DNA样本库,保护现有少数民族遗传资源基础上,利用维吾尔族基因组DNA样本库,通过现代分子生物学技术,对药物代谢酶基因CYP2D6在维吾尔族人群中的多态性分布规律进行了检测,为今后对该基因在维吾尔族中的代谢活性差异研究提供强有力的科学依据。 [研究方法] 在中央民族大学新入学学生中,通过调查问卷,询问三代内民族情况的方法,筛选三代以内无外族通婚史的少数民族健康成年人,在知情同意的原则下,按照临床采血标准进行采血,通过非离心柱法提取全血基因组DNA。按照人类遗传资源标准编码统一编号,并通过琼脂糖凝胶电泳及紫外分光光度计测定,剔除断裂及降解的DNA,剔除纯度较低样本,超低温保存,建立了中国少数民族基因组DNA样本库。在此基础上,通过分段克隆的方法,克隆CYP2D6全基因及其9个外显子序列。通过直接测序法对各样本进行了序列测定,利用DNAMAN, Primer5.0等生物学软件,对测序结果进行了生物信息学分析。检测了CYP2D6编码序列在维吾尔族人群中的SNP及单倍体型分布频率。 [研究结果] 1.中国少数民族DNA样本库的建立已经建成的DNA样本库目前包括朝鲜族、蒙古族、傣族、哈萨克族、维族、苗族、回族、藏族、土家族、彝族、侗族、羌族、壮族、瑶族、德昂族、纳西族、锡伯族、白族、柯尔克孜满族、哈尼族、布依族、仡佬族、畲族、黎族、水族、阿昌族、汉族、乌孜别克族、裕固族等30多个民族的765个基因组DNA样本。 2.CYP2D6在中国维吾尔族人群中的多态性分布我们在中国维吾尔族人群中筛查到13个未报导的等位基因型和2个已报导的等位基因型。13个未报导的等位基因的在中国维吾尔族人群中的分布频率介于5%到10%之间。已报导的等位基因分别是CYP2D6*2和CYP2D610D,它们的分布频率分别为10%和15%。 [结论] 1.初步建成中国少数民族基因组DNA样本库。 2.CYP2D6基因在中国维吾尔族人群的多态性分布频率与该基因在汉族及已报导的其他民族中的分布特点存在显著差异。
[Abstract]:[Purpose and significance] With the development of China's social and political and economic development, the integration of all ethnic groups is increasing, and the genetic resources of the pure ethnic minorities are becoming more and more Precious. In particular, the outflow of the genetic resources of the minority nationalities is out of the way, and it is urgent to increase the protection of the genetic resources of the minority nationalities Therefore, we have set up a library of genomic DNA samples containing a large sample of 56 national genetic information in China to fully preserve and protect the existing genetic Resources. These genetic resources can provide information on the origin, evolution and migration of the human race, and it is very important to conduct research on the dynamics of genetic factors and the dynamics of drug metabolism, even some of the mechanisms of genetic disease In recent years, the research of genetic pharmacology has become a hot spot in the field of medicine, especially between the genetic factors and the occurrence of adverse drug reactions, and all the countries of the world have given different degrees. Attention and attention. The existence of the polymorphism of the drug metabolizing enzyme is the drug effect and the adverse reaction. The main cause of the body difference is that all ethnic groups in our country have their own unique genetic characteristics, which will lead to different curative effects of the same drug in different ethnic groups. There is even an adverse reaction. Therefore, it is understood that the difference in the drug metabolism level of each ethnic group is to optimize the treatment dose, reduce the adverse reaction of the drug, and promote the traditional medicine of the ethnic minority. One of the key factors in the study. The analysis shows that the adverse reaction caused by the polymorphism of the P450 The total number of adverse reactions is 86%. Cytochrome P4502D6 (CYP2D6) is an important one of the most polymorphic drug metabolizing enzymes, metabolizes about 20-30% of the drug in clinic, and the drug reaction and drug caused by the polymorphism of CYP2D6 The main purpose of this study is to establish a library of genomic DNA of ethnic minorities in China, to protect the genetic resources of the existing minorities, and to use the DNA sample of the Uighur genome. By means of the modern molecular biology technology, the polymorphism distribution of the drug metabolizing enzyme gene CYP2D6 in the Uighur population is detected, and the metabolic activity of the gene in the Uygur population in the future is provided. differential research and supply A strong scientific basis.[Study Method] In the newly-enrolled students of the Central National University, through the questionnaire, the method of the three-generation internal-ethnic situation is asked, and the minority healthy adults who have no history of intermarriage within three generations are screened. Under the principle of consent, according to the clinical blood collection standard the whole blood genomic DNA is extracted by a non-centrifugal column method, the whole blood genomic DNA is coded according to the human genetic resource standard, In this paper, the genomic DNA sample library of China's national minorities is established. Methods The whole gene of CYP2D6 and its 9 exons were cloned. The samples were sequenced by direct sequencing, and the DNA MAN and Primer5 were used. Bioinformatics analysis of the results of the sequencing was carried out by the biological software such as 0. The coding of CYP2D6 was detected. The sequence is in the Uighur. SNPs in the family and the frequency of the haplotype distribution[Study Results] 1. The DNA sample library of China's national minority DNA sample library has been built, including Korean, Mongolian, Dai, Kazak and Uighur., the Miao, the Hui, the Tibetan, the Tujia, the Yi, the Dong, the Qiang, the Zhuang, the Chinese, the German, the Naxi, the Sibo, the Bai, the Kirk, the Manchu, the Hani, the Buyi, the family, the family, the family, the Li, the water, the Achang, 765 of more than 30 nationalities, such as the Han, the Uzbek, the Yugu, etc. Genomic DNA sample. 2. The polymorphism of CYP2D6 in the Chinese Uighur population. We screened 13 unreported allelic and 2 reported alleles in the Chinese Uighur population. genotype The distribution of 13 unreported alleles in the Chinese Uighur population ranged from 5% to 10%. The reported allele scores Don't be CY P2D6 * 2 and CYP2D610D, their distribution The frequency is 10% and 15%, respectively.[Conclusion] 1. The genomic DNA sample library of China's national minorities is preliminarily constructed. 2. CYP2D6
【学位授予单位】：中央民族大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R394

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