[Establishment of the cell line K562 with stable expression of hermap and hermap-siRNA].
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.
Department of Hematology and Oncology, Guangzhou Women and Children Medical Center, Guangzhou 510623, Guangdong Prvoince, China.
Yan-Mei
YM
Li
Sai-Jun
SJ
Gao
Tie-Zhen
TZ
Ye
Ying-Yi
YY
He
D004912:Erythrocytes / Q000737:chemistry; D020868:Gene Silencing; D006801:Humans; D020014:K562 Cells; D008565:Membrane Proteins / Q000235:genetics*; D010957:Plasmids; D034741:RNA, Small Interfering; D014162:Transfection
D008565:Membrane Proteins; D034741:RNA, Small Interfering