Objective: Constructing a plasmid containing tRNAVal promoter to express shRNA which mediates RNA interference.
Methods: 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.
Results: pUC-tRNAVallucRi suppressed the luciferase expression from pMAMneoLuc by 97.9%-9.5%.
Conclusion: The results showed that the tRNAVal shRNA plasmid could efficiently suppress luciferase expression in BHK-21.
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