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[Construction and application of inducible artificial microRNA expression vector targeting eIF3g gene].
- Source :
-
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology [Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi] 2011 Feb; Vol. 27 (2), pp. 135-8. - Publication Year :
- 2011
-
Abstract
- Aim: To construct an inducible artificial microRNA expression vector targeting eIF3g gene and use it to inhibit the expression of eIF3g in K562 cells.<br />Methods: The microRNA targeting human eIF3g was designed and obtained by PCR. After confirmed by sequencing, the microRNA was cloned into the pRevTRE2 plasmid. The Tet-off plasmids were transfected into K562 cells and selected for the stable tetracycline inducible K562/Tet-off transfectants. The pRevTRE2-eIF3g-miRNA plasmid was then transfected into stable K562/Tet-off transfectants and the expression of eIF3g was detected by Western blot.<br />Results: The microRNA targeting eIF3g was confirmed by sequencing. GFP fluorescence assay showed that the stable transfectants of Tet-off plasmids were under tetracycline control. Western blot results showed that the stable transfectants of pRevTRE2-eIF3g-microRNA inhibited eIF3g expression under the regulation of tetracycline.<br />Conclusion: The tetracycline-inducible artificial microRNA expression vector targeting eIF3g gene has been successfully constructed and effectively inhibits eIF3g expression in K562 cells.
- Subjects :
- Gene Expression Regulation, Leukemic drug effects
Gene Expression Regulation, Leukemic genetics
Humans
K562 Cells
Protein Synthesis Inhibitors pharmacology
Tetracycline pharmacology
Eukaryotic Initiation Factor-3 genetics
Eukaryotic Initiation Factor-3 metabolism
Genetic Vectors genetics
MicroRNAs genetics
Subjects
Details
- Language :
- Chinese
- ISSN :
- 1007-8738
- Volume :
- 27
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
- Publication Type :
- Academic Journal
- Accession number :
- 21315039