应用DHPLC检测多巴胺D2受体基因在中国人群中的多态性

通过变性高效液相色谱 (DHPLC)和DNA测序在 4 1个中国汉族人DNA样本中检测DRD2基因编码区和拼接区的单核苷酸多态性 (SNP) ,结果发现 3个SNP :Intron5的 2 77G/A、Exon7的 4 2C/T和Exon7的 1 2 9T/C .Intron5 2 77G/A是在中国汉族人群中发现的新SNP ,Exon7 4 2C/T和 1 2 9T/C在NCBIdbSNP中已有相应记录 (分别为rs4 986 92 1和rs6 2 75) ,它们均导致DRD2基因的同义突变 ,其中Exon7 1 2 9C等位基因频率在研究样本中高达 4 3.9% .这些结果为在中国汉族人群中开展DRD2基因相关的群体遗传学研究提供了遗传标记 .另外 ,还探讨了DHPLC检测突变的干扰因素及控制措施 ,为国内同行开展类似工作提供参考

Mutation Detection of the DRD2 Gene in Chinese Population by Denaturing High Performance Liquid Chromatography (DHPLC) and DNA sequencing

To provide available genetic polymorphisms for population genetics studies related to human dopamine receptor D2 (DRD2) gene in Chinese population,we used denaturing high performance liquid chromatography (DHPLC) and DNA sequencing to screen mutation in the coding and mRNA splicing regions of the DRD2 gene in 41 subjects of Han Chinese.We found three single-nucleotide polymorphisms (SNP): one is 277G/A in intron 5, the other two are 42C/T and 129T/C in exon 7.It is the first time for the 277G/A polymorphism in intron 5 of the DRD2 gene to be detected in Chinese Han population in present study.The 42C/T and 129T/C, both of which cause synonymous mutation in the DRD2 gene, had corresponding ID with rs4986921 and rs6275 respectively in NCBI dbSNP.We also genotyped the 277G/A polymorphism and the 129T/C polymorphism with DHPLC and restriction fragment length polymorphism (RFLP) respectively.The 277G/A had medium mutation frequency with 6.1% for 277A allele in our sample.The 129T/C was highly polymorphic (43.9% for 129C allele) in our population.Because we used healthy and randomly selected subjects from our study area, our results can be expected to stand for the situation of general Han Chinese of that area.For further studies on genetic etiology of some neurological diseases in Chinese population, present study provided some preliminary information of the DRD2 gene as a candidate susceptibility gene of those diseases.Additionally, we also assessed some interfering factors and their control methods during DHPLC analysis, which will be useful for our peers to do the similar research.