前列腺癌与前列腺增生相关基因的选择和克隆

目的 选择和克隆人前列腺癌(PCa)和前列腺增生(BPH)的相关基因。方法 根据当前前列腺癌与前列腺增生临床常规使用的肿瘤标记物和国内外基因表达谱的研究成果，选定了7个PCa上调表达。2个PCa下调表达基因和1个内参基因，采用聚合酶链反应(PCR)从正常人外周血DNA中扩增出上述10个基因的探针片段，并克隆至pGEM-T质粒中。重组质粒经PCR鉴定、EcoR Ⅰ单酶切或PSt Ⅰ和Nco Ⅰ双酶切鉴定，产物电泳检测。结果 检测结果表明已经成功地将10个基因的探针序列克隆入pGEM-T质粒载体中。结论 前列腺癌和前列腺增生相关基因质粒构建与探针制备为临床前列腺癌的血清学诊断及与前列腺增生症的鉴别提供了实验基础。

Selection and cloning of prostate cancer and BPH related genes

Objective To select the main related genes to prostate cancer (Pca) and BPH,then clone these genes into plasmid for further research.Methods Seven up-regulated genes,2 down-regulated genes and 1 internal control gene were selected according to clinically routine biomarker for PCa and current research result of gene expression about PCa and BPH.Using PCR technique,10 gene products were amplified from DNA extracted from normal human blood.Recombinants were acquired by cloning these genes' fragments into pGEM-T easy vector.Results Recombinants for 10 genes were validated by gel electrophoresis of insertion PCR amplicons and plasmid DNA fragments cleaved by EcoR I or by PSt I and Nco I.Conclusion The plasmid construction and probe preparation of related genes of PCa and BPH laid a basis for further research on serum diagnosis of PCa and BPH.