Linkage analysis between microsatellite markers and cleft lip and/or palate in Japanese families

我们研究了候选基因，BCL 3和附近的基因在19号染色体上，和非综合征性唇腭裂（CL/P）在日本家庭使用参数的方法。通过新泻大学牙科医院口腔颌面外科第二部门确定了9个多代CL/P家庭。在获得知情同意后，从60名个体中抽取血液样本，其中20人受到影响，并提取基因组DNA。用位于19q13.2的4个微卫星标记D19 S178、BCL 3、007/008和AC 1/AC 2进行PCR扩增，扩增产物经8%聚丙烯酰胺凝胶电泳分离，银染显示。此外，等位基因频率和杂合性值的四个标记进行了分析，为50个健康的无关的日本人。使用LINKAGE软件包的MLINK程序进行两点连锁分析，并计算每个家族的LOD评分，假设常染色体显性遗传模型具有降低的遗传率。我们还测试了连锁，包括表型信息，仅为每个家庭中的受影响的个人。因此，没有证据表明，在四个基因座的连锁仅受影响的模型。假定受影响个体的遗传距离为0.8、0.6和0.3，则在D19 S178、007/008和AC 1/AC 2位点两侧约1cM范围内未检测到连锁的证据。另一方面，当重组率设定为0.999时，BCL 3位点的最大LOD得分为0.206，这一结果表明与候选基因的连锁未被确定。

We examined linkage between candidate genes, BCL3 and nearby genes on chromosome 19, and non-syndromic cleft lip with or without cleft palate (CL/P) in Japanese families using parametric method. Nine multigenerational CL/P families were ascertained through the Second Department of Oral and Maxillofacial Surgery, Niigata University Dental Hospital. After informed consent was obtained, blood samples were drawn from 60 individuals, 20 of whom were affected, and genomic DNAs were extracted. PCR-amplified products using four microsatellite markers, D19S178, BCL3, 007/008 and AC1/AC2 located in 19q13.2, were separated by 8% polyacrylamide gel electrophoresis and visualized by silver staining. In addition, allele frequencies and heterozygosity values of four markers were analyzed for 50 healthy unrelative Japanese individuals. Two-point linkage analysis was carried out using the MLINK program of the LINKAGE package, and LOD scores were calculated for each family, assuming an autosomal dominant model of inheritance with reduced penetrance. We also tested for linkage including phenotypic information only for affected individuals in each family. Consequently, no evidence of linkage was detected at four loci in affected-only model. And assuming that penetrance for affected individuals was set alternatively at 0.8, 0.6, and 0.3, no evidence of linkage was detected within about 1 cM on both sides of D19S178, 007/008 and AC1/AC2 loci. On the other hand, the maximum LOD score for BCL3 locus was 0.206 at recombination fraction of 0 when penetrance was set at 0.999, this result indicated that linkage to candidate genes was not defined.