线粒体DNA编码区单核苷酸多态性研究
发布时间：2018-11-07 19:16
【摘要】：目的:筛选线粒体DNA(mtDNA)编码区频率较高的单核苷酸多态性(SNP)位点,研究其在中国汉族人群中的多态性;建立变性高效液相色谱(dHPLC)技术用于研究mtDNA 编码区SNP 的方法;利用所筛选的SNP 为线粒体DNA生物芯片的研究提供信息。 方法:针对文献报道的线粒体DNA 编码区多态性较高的区域,分成四段,即nt8162～8483(mtco1)、nt13070～13299(mtco2)、nt10287～10679(mtco3)、nt8507～8805(mtco4),分别设计引物,并且优化引物扩增条件,使之能同条件扩增;应用直接测序技术研究100 例中国汉族样本的mtco1、mtco2 多态性;随机抽取15 例样本,同时用测序和dHPLC 技术分别检测mtco3、mtco4 的多态性,在此基础上比较dHPLC 和测序两种方法结果的一致性,从而建立dHPLC 技术用于研究mtco3、mtco4 多态性的方法,再用已经建立的dHPLC 方法检测其余85 例样本的mtco3、mtco4 多态性。 结果:在100 例中国汉族人群中,mtco1、mtco2 共检出21 种变异,24种单倍型,基因多样性h 值为75.11%,偶合概率P 值为25.64%,其中,mtco1序列有15 例样本(15%)在COⅡ/tRNALys之间发生9bp(CCCCCTCTA)缺失,1 例样本发生9bp(CCCCCTCTA)插入;mtco3、mtco4 共检出23 个SNP 位点,23 种单倍型,基因多样性h 值为84.14%,偶合概率P 值为16.70%。四段序列联合起来,共检出44个变异位点,42种单倍型,基因多样性h值为94.79%,偶合概率P 值为6.16%。在所检测到的SNP 位点中,nt10400 及nt8701 的突变频率最高,均为57%,nt10398 为38%,nt8584 为20%,nt8414 为19%。 结论:只有不断扩大mtDNA 的检测范围才能提高其个体识别能力,满足法医学鉴定的需要;建立的dHPLC 方法可用于快速、准确地检测mtDNA编码区多态性,而且与直接测序法结果完全一致。
[Abstract]:Objective: to screen the single nucleotide polymorphism (SNP) loci with high frequency of mitochondrial DNA (mtDNA) coding region, and to study its polymorphism in Chinese Han population, and to establish a denaturing high performance liquid chromatography (dHPLC) technique for the study of SNP in mtDNA coding region. The selected SNP was used to provide information for the study of mitochondrial DNA biochip. Methods: the regions with high polymorphism of mitochondrial DNA were divided into four segments: nt8162~8483 (mtco1), nt13070~13299 (mtco2), nt10287~10679 (mtco3), nt8507~8805 (mtco4). The conditions of primer amplification were optimized so that the primers could be amplified under the same conditions. The mtco1,mtco2 polymorphism of 100 Chinese Han nationality samples was studied by direct sequencing technique. Fifteen samples were randomly selected and the polymorphism of mtco3,mtco4 was detected by sequencing and dHPLC respectively. The results of two methods, dHPLC and sequencing, were compared on the basis of which the method of using dHPLC to study mtco3,mtco4 polymorphism was established. The established dHPLC method was used to detect the mtco3,mtco4 polymorphism in the remaining 85 samples. Results: in 100 Chinese Han population, 21 variants and 24 haplotypes were detected by mtco1,mtco2. The gene diversity h value was 75.11, and the probability of coupling was 25.64. In 15 samples (15%) of mtco1 sequence, 9bp (CCCCCTCTA) deletion occurred between CO 鈪，
本文编号：2317323
[Abstract]:Objective: to screen the single nucleotide polymorphism (SNP) loci with high frequency of mitochondrial DNA (mtDNA) coding region, and to study its polymorphism in Chinese Han population, and to establish a denaturing high performance liquid chromatography (dHPLC) technique for the study of SNP in mtDNA coding region. The selected SNP was used to provide information for the study of mitochondrial DNA biochip. Methods: the regions with high polymorphism of mitochondrial DNA were divided into four segments: nt8162~8483 (mtco1), nt13070~13299 (mtco2), nt10287~10679 (mtco3), nt8507~8805 (mtco4). The conditions of primer amplification were optimized so that the primers could be amplified under the same conditions. The mtco1,mtco2 polymorphism of 100 Chinese Han nationality samples was studied by direct sequencing technique. Fifteen samples were randomly selected and the polymorphism of mtco3,mtco4 was detected by sequencing and dHPLC respectively. The results of two methods, dHPLC and sequencing, were compared on the basis of which the method of using dHPLC to study mtco3,mtco4 polymorphism was established. The established dHPLC method was used to detect the mtco3,mtco4 polymorphism in the remaining 85 samples. Results: in 100 Chinese Han population, 21 variants and 24 haplotypes were detected by mtco1,mtco2. The gene diversity h value was 75.11, and the probability of coupling was 25.64. In 15 samples (15%) of mtco1 sequence, 9bp (CCCCCTCTA) deletion occurred between CO 鈪，
本文编号：2317323
本文链接：https://www.wllwen.com/shekelunwen/gongan/2317323.html
