生物工程学报
生物工程學報
생물공정학보
CHINESE JOURNAL OF BIOTECHNOLOGY
2000年
3期
320-323
,共4页
赵新燕%蔡静莉%戴卫列%周伟国%李昌本%赵寿元
趙新燕%蔡靜莉%戴衛列%週偉國%李昌本%趙壽元
조신연%채정리%대위렬%주위국%리창본%조수원
人促血小板生成素受体c-Mpl%RT-PCR%转染%K562细胞表达
人促血小闆生成素受體c-Mpl%RT-PCR%轉染%K562細胞錶達
인촉혈소판생성소수체c-Mpl%RT-PCR%전염%K562세포표체
Human thrombopoietin receptor c-Mpl%RT-PCR%transfection%K562 cell%expression
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。
以人HEL細胞總RNA為模闆,採用RT-PCR方法擴增瞭人促血小闆生成素受體c-Mpl編碼區全長1.9kb cDNA,測序結果錶明與已報道的序列一緻。然後構建瞭c-mpl的pcDNA3錶達載體pcMPL,轉染不錶達c-mpl的K562細胞後,經G418抗性篩選,Northern blot和Southern blot檢測證實穫得穩定錶達c-mpl的細胞株。為進一步研究c-Mpl的生物學功能提供有用的實驗材料。
이인HEL세포총RNA위모판,채용RT-PCR방법확증료인촉혈소판생성소수체c-Mpl편마구전장1.9kb cDNA,측서결과표명여이보도적서렬일치。연후구건료c-mpl적pcDNA3표체재체pcMPL,전염불표체c-mpl적K562세포후,경G418항성사선,Northern blot화Southern blot검측증실획득은정표체c-mpl적세포주。위진일보연구c-Mpl적생물학공능제공유용적실험재료。
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.