Construction of a tRNAVal promoter plasmid expressing shRNAs in mammalian cells which mediates RNA interference / 中华实验和临床病毒学杂志

ABSTRACT

<p><b>OBJECTIVE</b>Constructing a plasmid containing tRNAVal promoter to express shRNA which mediates RNA interference.</p><p><b>METHODS</b>A tRNAVal gene was amplified from human genomic DNA by PCR and replaced the last several bases of 3' end by a linker. The tRNAVal promoter after artificial mutation followed a shRNA sequence to luciferase was cloned into pUC18, Puc-tRNAVal, lucRi Cotransfected with pMAMneoLuc into BHK-21 cell to detect the effect of luciferase expression.</p><p><b>RESULTS</b>pUC-tRNAVallucRi suppressed the luciferase expression from pMAMneoLuc by 97.9%-9.5%.</p><p><b>CONCLUSION</b>The results showed that the tRNAVal shRNA plasmid could efficiently suppress luciferase expression in BHK-21.</p>