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  • Title: [Constructing recombinant plasmid with prostate-specific membrane antigen promoter and reverse enhancer and comparing the transcription activity of enhancers in different directions].
    Author: Zeng H, Liao YC, Wang F, Wu Q, Yang YR.
    Journal: Sichuan Da Xue Xue Bao Yi Xue Ban; 2005 Nov; 36(6):782-5. PubMed ID: 16334552.
    Abstract:
    OBJECTIVE: To assess the specificity of prostate-specific membrane antigen(PSMA) promoter and enhancer in controlling gene expression and to compare the activity of enhancers in different directions for choosing the most suitable prostate-specific PSMA controlling elements. METHODS: PSMA enhancer gene was amplified with PCR, then the enhancer gene was subcloned into the expressing vector pEGFP-PSMA(Pro) reversely to construct the recombinant plasmid pEGFP-PSMA(E(r)-p), which was transfected into different cell lines such as LNCaP, PC-3,MCF-7,A549. Green fluorescent protein (GFP) expression was observed and compared to other recombinants constructed previously. RESULTS: The recombinant plasmid with reverse enhancer was successfully constructed, the PSMA promoter and enhancer showed modulating activity in PSMA-expressed cell line uniquely. PSMA enhancer could increase 30-fold transcriptional activity over the basal level achieved by PSMA promoter alone, and no impact of the direction on the activity of enhancer was noted. CONCLUSION: PSMA promoter/ enhancer is specific to PSMA-expressed cells. The transcriptional activity of reverse enhancer is similar to that of enhancer. PSMA promoter/enhancer has the potential for use in targeted gene therapy of prostate adenocarcinoma.
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