[Cloning of a full length cDNA of human thrombopoietin receptor c-Mpl and construction of engineered cells that stably express c-mpl].
A full length cDNA fragment encoding for human thrombopoietin receptor c-Mpl has been amplified by RT-PCR from the total RNA of human HEL cells. The complete sequence of the cloned cDNA was determined and is identical to that previously reported. Then the fragment was subcloned into the mammalian expression vector pcDNA3 and the resulting plasmid is designated as pcMPL. K562 cells, which do not express c-mpl, were transfected with pcMPL and pcDNA3, respectively. The transformants were selected with G418 and then tested by Northern and Southern blotting. A group of engineered cell lines stably expressing c-mpl have been obtained, which will facilitate further research on the signaling mediated by c-Mpl.
Zhao
X Y
XY
State Key Laboratory of Genetic Engineering, Fudan University, Shanghai.
Cai
J L
JL
Dai
W L
WL
Zhou
W G
WG
Li
C B
CB
Zhao
S Y
SY
D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry*; D006801:Humans; D020014:K562 Cells; D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D018121:Receptors, Cytokine; D053628:Receptors, Thrombopoietin; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection
D018076:DNA, Complementary; D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins; D018121:Receptors, Cytokine; D053628:Receptors, Thrombopoietin; C082218:MPL protein, human