IDENTIFICATION OF SINGLE NUCLEOTIDE POLYMORPHISMS IN CANCER-RELATED GENES

单核苷酸多态性（SNPs）代表了理解复杂疾病的遗传标记的丰富和有用的来源。生物学上重要基因编码区的SNPs（cSNPs）可能会在功能上改变蛋白质产物。我们研究了40个与癌症相关的基因的DNA序列变异性，包括癌基因、肿瘤抑制基因和参与细胞周期控制的基因。设计扩增每个外显子和内含子边界的引物对，并用于扩增来自四个欧洲裔美国人（CEPH家族父母）、四个非洲裔美国人和四个亚裔美国人样品的DNA。对产品的分析表明，超过90%的产品尺寸正确。通过变性高效液相色谱法（DHPLC）分析产物的变化。共检测到42个潜在变异，其中35例通过测序检测到SNP。为了确认这些变异体的分离，在整个CEPH家系中检查了其中几个。我们证明了变异体确实与侧翼微卫星标记沿着分离。我们总共扫描了约70 kb的序列，发现核苷酸多样性为2.5 X 10-5。这个数字与文献中报道的使用DNA芯片或DHPLC的其他基因的结果一致（1.7-3.1 X 10-5）。因此，DHPLC是一种有效的方法，以确定变异的基因，然后可以用作遗传标记，在人口为基础的研究。 - 癌症遗传学，人类遗传学，肿瘤抑制基因，-人体组织，液体，细胞等。

Single nucleotide polymorphisms (SNPs) represent an abundant and useful source of genetic markers to understand complex diseases. SNPs in coding regions (cSNPs) of biologically important genes are likely to functionally alter the protein product. We have investigated the variability in DNA sequence in 40 genes related to cancer including oncogenes, tumor suppressor genes, and genes involved in cell cycle control. Primer pairs amplifying each exon and intron boundary were designed and used to amplify DNA from four European Americans (CEPH family parents), four African Americans, and four Asian American samples. Analysis of the products showed that over 90% produced a correctly sized product. The products were analyzed for variation by denaturing high performance liquid chromatography (DHPLC). A total of 42 potential variants were detected and the SNP was detected by sequencing in 35 cases. To confirm the segregation of these variants several of them were examined in entire CEPH pedigrees. We demonstrated that the variants did segregate along with flanking microsatellite markers. In total we scanned about 70 kb of sequence and found a nucleotide diversity of 2.5 X 10-5. This figure agrees with that reported in the literature for other genes using either DNA chip or DHPLC (1.7-3.1 X 10-5). Therefore DHPLC is an effective method to identify variations in genes that can then be used as genetic markers in population based studies. - Cancer genetics, Human Genetics, Tumor suppressor genes, - Human Tissues, Fluids, Cells, etc.