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[Screening differentially expressed genes in denucleated K562 cells with restriction display technique]
Min Wei1, Wen-li Ma, Yan-bin Song
1Institute of Molecular Biology, Southern Medical University, Guangzhou 510515, China. phenney@fimmu.com
Objective:
To screen differentially expressed genes in cytochalasin B (CB)-induced denucleated K562 cells by restriction display (RD) technique.
Methods:
The total RNA was isolated and purified from K562 cells before and after CB (10 mug/ml) treatment. The mRNA from both treated and untreated K562 cells were reversely transcribed into cDNA, and the differentially expressed genes were separated using RD technique combined with polyacrylamide gel electrophoresis and sliver staining, followed by cloning, sequencing and homology analysis against GenBank database of these genes.
Results:
Seven differentially expressed genes were identified in CB-treated cells including aquaporin 1 (AQP1) gene, which was verified to be up-regulated after CB treatment by RT-PCR.
Conclusion:
AQP1 gene might be in close association with the regulation of denucleation processes and CB-induced proliferation inhibition of K562 cells.
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