鸟苷酰环化酶C质粒标准品的构建  

Construction of the plasmid DNA of guanylyl cyclase-C

作　　者：王唯一[1] 毛振彪[1] 徐政府[1] 俞智华[1] 周正斌[1] 张冬雷[1] 

机构地区：[1]南通大学附属医院消化内科,江苏226001

出　　处：《交通医学》2006年第5期503-504,共2页Medical Journal of Communications

摘　　要：目的:制备人鸟苷酰环化酶C(GC-C)质粒标准品,为实时荧光定量PCR检测GC-C含量,探讨大肠癌患者外周血单个核细胞(PBMC)GC-C mRNA表达水平及临床意义奠定基础。方法:在鸟苷酰环化酶C(GC-C)基因高保守区设计特异性的引物和探针,RT-PCR扩增目的片段并连接于pGEM-T载体,以得到GC-C质粒标准品。结果:经过蓝白斑筛选,PCR或EcoR I酶切初步鉴定,阳性克隆测序分析确定GC-C质粒标准品正确性。结论:正确构建GC-C质粒标准品为RFQ-PCR标准曲线的绘制及GC-C的定量检测奠定了基础。Objectives：To Construct the plasmid DNA of guanylyl cyclase-C,and make a foundation for measurement of the expression level of guanylyl cyclase-C （GC-C）in the Peripheral blood mononuclear cell （PBMC）and elucidation of the clinical significance of its level in colorectal cancer patients by establishing real-time fluorenscence quantitative polymerase chain reaction （RFQ-PCR）,Methods：Specific primers and TaqMan probe were designed,and GC-C fragments were amplified by RT-PCR and connected with pGEM-T to get plasmid DNA of GC-C.Results：positive clone was assayed correctly by detection of the sequence. Conclnsions：The plasmid DNA of GC-C constructed correctly made a foundation for the creation of the standard curve and quantitative detection of GC-C.

关 键 词：鸟苷酰环化酶C 结肠直肠肿瘤 实时荧光定量 

分 类 号：R730.54[医药卫生—肿瘤]