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目的研究先天性髋关节脱位(CDH)与X染色体之间有无连锁关系,为进一步寻找疾病易感基因的全基因组扫描进行初步尝试。方法以4个云南地区CDH家系为研究对象,提取DNA。选用人类连锁协作中心(The Cooperative Human linkage Center,CHLC)开发的17个分布于X染色体的微卫星多态标记进行PCR扩增,产物进行非变性凝胶电泳,银染色。凝胶分析系统行等位基因分型。应用LINKAGE软件包在不同的重组率条件下行参数型连锁分析,设疾病基因频率为0.02。结果所有位点表现了较高杂合度和多态性,基因型数据符合孟德尔定律。在X染色体遗传标记DXS8091连锁分析时,发现重组率=0.0时,Lod=2.184339,提示可能存在连锁。又在DXS8091附近每隔3cm的范围内选择了7个多态性位点进行同样的实验,但两点连锁Lod<1,不支持存在两点间的连锁。提示家系CDH与各微卫星位点不存在连锁关系。结论排除云南地区CDH家系致病基因位于X染色体。
Objective To study whether there is a linkage between congenital dislocation of the hip (CDH) and the X chromosome, and to conduct a preliminary attempt to further investigate the genome-wide scanning of disease-susceptible genes. Methods Four CDH pedigrees in Yunnan were used as research objects to extract DNA. Seventeen SSR markers distributed on the X chromosome were selected from the Cooperative Human Link Center (CHLC) for PCR amplification. The products were subjected to non-denaturing gel electrophoresis and silver staining. Gel analysis system line allele typing. Using LINKAGE package under different recombination rates, a parametric linkage analysis was carried out. The frequency of disease genes was 0.02. Results All sites showed higher heterozygosity and polymorphism, and genotype data accorded with Mendel’s law. In the linkage analysis of the X chromosome genetic marker DXS8091, it was found that Lod = 2.184339 at the recombination rate = 0.0, indicating that there may be a linkage. In the vicinity of DXS8091, 7 polymorphic sites were selected every 3 cm for the same experiment, but the two-point linkage Lod <1 does not support the existence of a linkage between two points. It is suggested that there is no linkage between family CDH and microsatellite loci. Conclusion It is excluded that the CDH pedigree in Yunnan Province is located on the X chromosome.