cDNA cloning and function analysis of two novel erythroid differentiation related genes |
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Authors: | Xin Wang Duncheng Wang Xing Chen Meiru Hu Jian’an Wang Yan Li Ning Guo Beifen Shen |
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Institution: | (1) Beijing Institute of Basic Medical Sciences, 100850 Beijing, China |
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Abstract: | Our previous studies showed that some nuclear proteins that were expressed especially during terminal differentiation of erythroid
cells might interact directly or indirectly with HS2 sequence to form the HS2-protein complexes and thus play an important
role in the globin gene regulation and erythroid differentiation. Monoclonal antibodies against the nuclear proteins of terminal
differentiated erythroid cells, including intermediate and late erythroblasts of human fetal liver and hemin induced K562
cells, were prepared by hybridoma technique. The monoclonal antibodies were used to screen λ-gtll human cDNA expression library
of fetal liver in order to obtain the relevant cDNA clones. By the analysis of their cDNA clones and the identification of
the proteins’ functions, the regulation mechanism of the HS2 binding proteins might be better understood. Two cDNA clones
(GenBank accession number AF040247 and AF040248 respectively) were obtained and one of them owns a full length and the other
encodes a protein characterized by a leucine-zipper domain. Both of them were expressed differentially in K562 cells and hemin-induced
K562 cells. The evidence suggested that both of them were involved in erythroid differentiation. We investigated the expression
pattern ofEDRF1 andEDRF2 by RT-PCR technique. The results of RT-PCR suggested that EDRF1 and EDRF2 might play a critical role in early stage of organ
development and histological differentiation. EDRF1 and EDRF2 might start the program of erythroid development, and also regulate
the development of erythroid tissue and the expression of globin gene at different stage of the development. |
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Keywords: | erythroid differentiation HS2 cDNA cloning leucine zipper domain RT-PCR |
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