Analysis of the effect of pcDNA3.1 on house keeping gene expression
- VernacularTitle:pcDNA3.1载体对内参基因表达影响分析
- Author:
Ke CHEN
- Publication Type:Journal Article
- Keywords:
pcDNA3.1;
House keeping Gene;
Transfection;
Transcription;
Expression
- From:
Journal of Chongqing Medical University
2003;0(06):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of pcDNA3.1 vector on house keeping gene expression and set guide to further study work in using pcDNA3.1. Methods:5 cell lines were transfected with pcDNA3.1,48 h post transfection total RNA was extracted and applied for real time PCR to detect the RNA level of GAPDH、 ?-actin、18sRNA difference.Meanwhile,extract total protein and analyze the GAPDH and?-actin expression by Western blot.Results:RNA and protein levels all show of a decrease in 5 cell lines after transfection,with the most obviously ?-actin.Conclusion:pcDNA3.1 might have non specifinhibition effect to cell,when using pcDNA3. 1 for gene function research,it is not recommend to use ?- actin as reference gene.
