【摘要】 目的应用变性高效液相色谱(DHPLC)检测CD31 563位点单核苷酸多态性。方法聚合酶链反应(PCR)扩增健康正常人位于17q23编码CD31基因第8外显子563位点附近的DNA片段,应用DHPLC技术对扩增片段进行多态性分析,将不同峰型的PCR片段进行序列测定,并与参考序列对照分析。结果PCR扩增片段为203 bp,经DHPLC分析有3种不同峰型,经与测序结果比较, G／G纯合峰型、A／A纯合峰型、G／A杂合峰型明显不同,3种基因型的个体得到明确区分。74名健康正常人中CD31-563S(AGC)的基因频率为0．514,CD31-563N(AAC)的基因频率为0．486。结论DHPLC可以高效、经济、准确的检测CD31563位点单核苷酸多态性。更多还原

【Abstract】 Objective Detection of single nucleutide polymorphisms in CD31-563 codon by denaturing high-performance liquid chromatography(DHPLC). Methods The eighth exon fragments of CD31 code gene in chromosome 17q23 were amplified by PCR. The fragments were analysed with DHPLC and were sequenced and compared with the sequences available in National Center for Biotechnology Information ( NCBI) database. Results The length of the amplified fragments is 203bp. There are three kinds of pictures when the fragments are analysed by DHPLC, after comparing with direct sequencing, the pictures of G/G homozygous、A/A homozygous、G/A heterozygous are obviously different, thus the individuals with the three kinds of genetype can be distinguished accurately. In 74 healthy people studied,the gene frequencies of CD31-563S(AGC) is 0.514, the gene frequencies of CD31-563N( AAC) is 0. 486. Conclusion DHPLC can effectively, economically and accurately detect the single nucleutide polymorphisms in CD31 -563 codon.更多还原