Construction of an AFP enhancer-regulated siRNA expression plasmid vector

Abstract

Objective To construct an AFP enhancer-regulated siRNA expression plasmid vector.Methods The AFP gene was extracted from HePG2 cells, constructing plasmid PGenesil-AFP by orientation cloned.After DNA extraction and amplification, Plasrnid pGenesil-AFP was transformed into competent cell of DHS.Constructing plasmid vector pGenesil-AFP-shRNA to aim at the target gone sruvivin' s sequence.Results After p]asmid pGenesil-AFP-shRNA evaluated by gene sequencing analysis and enzyme cutting with both XbaⅠ and Hind Ⅲ, obtain a fragment of 856 bp and a fragment of 4506 bp.It is prove that Plasmid can be successfully constructed.Measured by ultraviolet spectrophotometer, the concentration of plasmid is 1.3 g/L and A value is 1.94.This shows that the purity of plasmid is high.Conclusion The successful construction of an AFP enhancer-regulated survivin shRNA integration plasmid. Key words: Carcinoma,hepatoceUular; AFP; Small interfering RNA; Plasmid vector