Construction of eukaryotic expression vector expressing double shRNA sections targeting Survivin gene in Bx-PC3 cells
- VernacularTitle:胰腺癌Bx-PC3细胞Survivin基因ShRNA质粒表达载体的构建
- Author:
He HUANG
;
Pei WU
;
Shujian HONG
;
Jiading MAO
;
Jing RUI
- Publication Type:Journal Article
- Keywords:
Survivin gene;
RNA interference;
small hairpin RNA;
pancreatic cancer cell
- From:
Chinese Journal of Clinical Pharmacology and Therapeutics
2002;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To construct eukaryotic expression vector expressing double shRNA sections targeting Survivin gene.METHODS: Eukaryotic expression vector expressing double shRNA sections targeting Survivin gene were designed and chemically synthesized.They were directionally inserted into plasmid pGenesil-1 with respectively U6 promoter and termination code,the common green fluorescence protein(EGFP) gene and Neo gene. In this way,the vector of pGenesil-1 shRNA containing 2 sections of Survivin shRNA were constructed and they were transfected into the pancreatic cancer cell Bx-PC3.Transfection was detected by fluorescence microscope.The inhibition expression of Survivin mRNA was measured by RT-PCR.RESULTS: HE1 and HE2 plasmids were identified by the biocatalyst cut which confirmed the exactitude and were analyzed by the sequence analysis which verified the perfect clone plasmid inserted by them.CONCLUSION: A eukaryotic expression vector of double short hairpin RNA for Survivin gene is successfully constructed.The pancreatic cancer cells Bx-PC3 succeed to be transfected and expression of Survivin mRNA is inhibited obviously.
