【摘要】 从前列腺癌细胞系 LNCa P中提取总 RNA,经 RT-PCR扩增前列腺特异性膜抗原 (PSM)靶序列 ,将其定向插入带 T7RNA聚合酶启动子的 p SPORT1质粒 ,转入大肠杆菌 DH5 α,经蓝白斑筛选获得重组质粒 .以此重组质粒线性化 DNA为模板 ,以地高辛标记的 r UTP及其他三种 r NTP为底物 ,经 T7RNA聚合酶体外转录 ,成功地制备了地高辛标记的 PSM特异性 RNA探针 ,为研究以 PSM为分子生物学指标的前列腺癌分期诊断奠定了基础 .更多还原

【Abstract】 Total RNA was extracted from prostate cancer cell line LNCaP, and prostate specific membrane antigen (PSM) gene target sequence was inserted into the pSPORT1 vector. Which possessed T7 RNA polymerase promoter. Recombined plasmid was transformed into E.coli DH5 α and chosen by "white blue plaque selection". The recombinant plasmid pSPORT1 PSM was identified by restricted enzyme map; PCR amplification and sequence analysis. The Digoxin labeled RNA probe was transcribed in vitro using the linear recombinant plasmid as template. It laid a foundation for researching the stage diagnosis of prostate cancer with the PSM as molecular biological marker.更多还原