Screening of proteins that interact with human thrombopoietin receptor c-Mpl using yeast two-hybrid system |
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Authors: | Xinyan Zhao Libing Feng Weiguo Zhou Weilie Dai Changben Li Shouyuan Zhao |
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Institution: | (1) State Key Laboratory of Genetic Engineering, Institute of Genetics, Fudan University, 200433 Shanghai, China |
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Abstract: | Thrombopoietin (TPO) is the major cytokine involved in platelet production and exerts its effects via the receptor c-Mpl. The yeast two-hybrid system has been used to screen the proteins interacting with c-Mpl. First, the cDNA fragment of c-Mpl intracellular domain was cloned into two-hybrid vector pAS2, and the resulting plasmid is designated as pASMM. Then a human placenta cDNA library was screened using the pASMM as a target plasmid. Seven positive clones were isolated from 150 000 independent transformants. Sequence analysis of one of the positive clones demonstrates that a part of coding sequence of vimentin from 611 bp to 3' end and flanking non-translation region was obtained. Therefore, there is an interaction between vimentin and TPO receptor. The results suggest that cytoskeletal protein may play an important role in TPO signal transduction pathway. |
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Keywords: | thrombopoietin receptor c-Mpl yeast two-hybrid system vimentin interaction |
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