title: [Construction and expression in vitro of an RU486 inducible vector carrying DsRed protein].
abstract: The regulation of a target gene expression is very important in gene therapy. However, constitutive or inappropriate expression of the genes with traditional expression system may interfere with the effect of the gene therapy, even may lead to lethal side effect. We constructed an RU486 inducible eukaryotic vector carrying DsRed protein and evaluated its regulatable effect in vitro. The single vector named PDC-RURED was constructed with molecular biological methods, which contained DsRed gene, promoter and RU486-inducible system. To minimize any potential interference, we spaced the two transcriptional elements with a 1.6 kb insulator. The vector was identified by different enzyme restrictions, sequencing analysis and PCR assay. We demonstrated the regulatable expression of this vector after transfection in HEK293 cells by fluorescence microscopy and flow cytometry. In the absence of RU486, no significant DsRed protein activation was observed, whereas in the presence of RU486 up to 40 fold activation of the DsRed protein was observed in cultured cells. The data show that the novel eukaryotic expression plasmid vector can be used to regulate the expression level of genes of interest in appropriate time under the control of RU486. This inducible expression vector provides a powerful tool for the research of gene regulation and gene therapy.
authors.affiliation: Department of Oncological Surgery, 81st Hospital of PLA, Nanjing 210002, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Chen
authors.forename: Xuchao
authors.initials: X
authors.lastname: Xue
authors.forename: Guoen
authors.initials: G
authors.lastname: Fang
authors.forename: Changqing
authors.initials: C
authors.lastname: Su
authors.forename: Qijun
authors.initials: Q
authors.lastname: Qian
mesh_terms: D002460:Cell Line; D005456:Fluorescent Dyes / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D007668:Kidney / Q000166:cytology; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000235:genetics; D015735:Mifepristone / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D014162:Transfection
chemical_list: D005456:Fluorescent Dyes; D008164:Luminescent Proteins; C413663:fluorescent protein 583; D015735:Mifepristone