关键词: 细胞株, 增强型绿色荧光蛋白, 前列腺肿瘤, 蛋白转导结构域, 前列腺特异抗原

Abstract:

Objective〓〖WTBZ〗We constructed an expression vector containing EGFP fused to the viral TAT transduction domain and driven by the prostate-specific antigen promoter (pPSA-TAT-EGFP) by gene cloning technology. The targeting expression of pPSA-TAT-EGFP was examined in prostate cancer cells and colon cells. 〖WTHZ〗Methods〓〖WTBZ〗The vector pEGFP-1 served as the framework, and the fragment with the TAT sequence and EGFP was produced by using polymerase chain reaction. After digestion with SalI and NotI enzymes, the fragment was inserted into the T cloning vector. The EGFP expression plasmid driven by the PSA promoter (pPSA-EGFP) was digested overnight with SalI and NotI, and a large fragment containing the PSA promoter was purified by gel purification. The TAT-EGFP, cut with SalI and NotI from the T-vector, was linked with the large fragment with the PSA promoter sequence and generated the pPSA-TAT-EGFP construct. The construct was confirmed by restriction enzyme digestion and DNA sequencing, and tested by transfection assay. 〖WTHZ〗Result〓〖WTBZ〗The expression vector pPSA-TAT-EGFP was constructed. It was expressed in prostate carcinoma cells, but was not detected in colon cells. 〖WTHZ〗Conclusion〓〖WTBZ〗A slightly higher expression was observed in cells transfected with EGFP fused to TAT compared to the EGFP control plasmid in prostate cancer cells, but no detectable expression was shown in colon cells.

Key words: Prostate-specific antigen, Protein transduction domain, Enhanced green fluorescent protein, Prostate neoplasms, Cell lines