Construction and identification of shRNA Ientivirus vector pIasmid of human SNCG gene
LI Wenyi
FAN Wenjuan
XU Hong
FAN Jiangtao
SUN Dan
Abstract：Objective To construct short hairpin RNA (shRNA)lentiviral vector plasmid of human SNCG gene and to detect the changes of SNCG mRNA and protein expression in human endometrial carcinoma cells HEC-1-A and Ishikawa after transfection.Methods Three SNCG gene specific shRNA sequences were designed,which were SNCG-KD1,SNCG-KD2 and SNCG-KD3,respectively.Using Lipofectamine 2000 to construct a lentiviral vector plasmid in 293T cells.Hu-man endometrial carcinoma HEC-1-A and Ishikawa cells were transfected with SNCG-KD1,SNCG-KD2 and SNCG-KD3 lentiviral vector plasmids.Using real-time PCR to detect the expression of SNCG mRNA,and Western blotting to detect SNCG protein expression.Results Age I and EcoR I were used to connect the three groups of positive transformation.The specific bands were in agreement with the predicted results by 1% agarose gel electrophoresis.The results of the analysis and identification of Lite Chromas (VERSION 2.01)gene sequencing were in agreement with GeneBank database align-ment,which showed that the synthesis of three pairs of shRNA Oligo DNA SNCG sequences SNCG-KD1,SNCG-KD2 and SNCG-KD3 inserted correctly.After the three lentiviral vectors were transfected into HEC-1-A and Ishikawa cells,the ex-pression of SNCG mRNA and protein in HEC-1-A and Ishikawa cells was significantly decreased (P <0.05).Conclusion Three shRNA SNCG lentiviral vector plasmids are successfully constructed and transfected into HEC-1-A and Ishikawa cells,which may effectively knock down the SNCG gene and inhibit the expression of SNCG mRNA and protein in HEC-1-A and Ishikawa cells.
Keywords：SNCG geneshort hairpin RNAlentivirus vector plasmidendometrial carcinomagene silencing
Publication Date：2016-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 13-16 )