[Construction and expression of RU486-inducible eukaryotic vector carrying red fluorescent protein].
OBJECTIVE
To construct an inducible eukaryotic vector carrying red fluorescent protein (DsRed) and evaluate the regulation of DsRed gene expression in vitro.


METHODS
The vector pRS17-RUDsRed containing DsRed gene, promoter and RU486-inducible system was constructed using molecular biological methods. To minimize potential interference, the two transcriptional elements were spaced with a 1.6 kb insulator. Fluorescence microscopy and flow cytometry were used to observe the activation of this regulatable vector after transfection in MFC cells.


RESULTS
The vector was identified by digestion with different restriction enzymes, sequencing and PCR. In the absence of RU486, the cells transfected with the vector exhibited very low DsRed protein expression, and the addition of RU486 induced efficient DsRed expression in the cells.


CONCLUSIONS
The RU486-inducible eukaryotic vector carrying DsRed protein allows effective regulation of the target gene expression in vitro, which provides a useful tool for gene regulation and gene therapy studies.
Chen
Jian
J
Department of Oncosurgery , 81 Hospital of PLA , Nanjing 210002, China. chenbond@126.com
Xue
Xu-chao
XC
Fang
Gao-en
GE
Su
Chang-qing
CQ
Qian
Qi-jun
QJ
D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D015735:Mifepristone / Q000494:pharmacology*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D013274:Stomach Neoplasms / Q000235:genetics* / Q000473:pathology; D014407:Tumor Cells, Cultured; D000097573:Red Fluorescent Protein
D008164:Luminescent Proteins; D015735:Mifepristone; D000097573:Red Fluorescent Protein