| Purpose:To construct the adenovirus mediated dual-luciferase reporters assay system that is stable and reliable to detect activities of promoters and can acquire highly repeatable results and to screen out 1 or 2 promoters that can perform high-efficiency expression.Experimental Design:Firstly, the coding genes of Renilla luciferase and firefly luciferase were constructed respectively into the different regions, ΔE1 and ΔE3, of the same adenoviral vector to obtain the dual-luciferase reporter assay system. After identification, amplification, purification and titration determination, the recombinant adenovirus were used to transfect 293, LO2, Huh7, HepG2, H460, H1299, Jurkat, K562, Hela, SK-OV-3 and primary muscle cell of mice to evaluate the feasibility and reliability of the adenovirus mediated dual-luciferase reporters assay system by detecting the activities of the promoters, finally, we investigated the performance of a series of promoters newly modified by this adenoviral system.Results:The adenovirus mediated dual-luciferase reporters assay system was successfully constructed. Performances of some promoters commonly used in adenoviral vector are consistent with previous studies. Successfully, CCAU and CCEF, two promoters that express efficiently, were obtained.Conclusions:The adenovirus mediated dual-luciferase reporters assay system as a new method of studying promoters is stable, reliable and accurate. |
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