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目的 :研究pc 1基因表达产物在细胞内的定位。方法 :采用巢式PCR技术 ,从原始克隆质粒中扩增出可编码蛋白分子的pc 1cDNA片段 ,分别将其克隆入含EGFP和MYC标签的真核表达载体中 ;用脂质体介导法将其转染入人前列腺癌细胞C4 2中 ;通过免疫细胞化学方法和激光共聚焦显微镜技术检测pc 1表达产物在细胞内的定位。结果与结论 :pc 1表达产物定位于细胞质中 ,为进一步研究其生理、生化功能奠定了基础。
Objective: To study the localization of pc 1 gene expression products in cells. METHODS: Nested PCR was used to amplify pc 1 cDNA fragments encoding protein molecules from the original clonal plasmids and cloned into eukaryotic expression vectors containing EGFP and MYC tags respectively. The liposome-mediated method was used. The transfected cells were transfected into human prostate cancer cells C4 2 and the localization of pc 1 expression products in the cells was detected by immunocytochemistry and laser confocal microscopy. RESULTS AND CONCLUSION: The pc1 expression product was localized in the cytoplasm, which laid the foundation for further study of its physiological and biochemical functions.