Establishment of the cell line K562 with stable expression of hermap and hermap-siRNA.
- Author:
Yan-Mei LI
1
;
Sai-Jun GAO
;
Tie-Zhen YE
;
Ying-Yi HE
Author Information
1. Department of Hematology and Oncology, Guangzhou Women and Children Medical Center, Guangzhou 510623, Guangdong Prvoince, China.
- Publication Type:Journal Article
- MeSH:
Erythrocytes;
chemistry;
Gene Silencing;
Humans;
K562 Cells;
Membrane Proteins;
genetics;
Plasmids;
RNA, Small Interfering;
Transfection
- From:
Journal of Experimental Hematology
2010;18(5):1306-1309
- CountryChina
- Language:Chinese
-
Abstract:
In order to establish K562 line with stable expressions of hermap and hermap-siRNA, amplified hermap and hermap-siRNA were cloned into pEGFP-c1 and pRNAT to acquire hermap-pEGFP-c1 and hermap-siRNA-pRNAT, respectively. These two plasmids were electrotransferred into K562 cells, then were followed by culturing with G418. The result showed that the transfer rate of hermap-pEGFP-c1-K562 and hermap-siRNA-pRNAT-K562 plasmids were 10.0% and 9.3%, respectively. After selective culture by G418, these two cell lines were still able to express GFP. It is concluded that the eukaryotic expression plasmids containing hermap and hermap-siRNA have been successfully constructed, and the cell lines of hermap-K562 and hermap-siRNA-K562 are established, definitely contributing to further functional investigation on HERMAP and its interaction with other proteins.
