以人HEL细胞总RNA为模板，采用RT-PCR方法扩增了人促血小板生成素受体c-Mpl编码区全长1.9kb cDNA，测序结果表明与已报道的序列一致。然后构建了c-mpl的pcDNA3表达载体pcMPL，转染不表达c-mpl的K562细胞后，经G418抗性筛选，Northern blot和Southern blot检测证实获得稳定表达c-mpl的细胞株。为进一步研究c-Mpl的生物学功能提供有用的实验材料。%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.
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