Expression cloning of oligomerization-activated genes with cell-proliferating potency by pseudotype retrovirus vector |
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Authors: | Abe Akihiro Emi Nobuhiko Kanie Tadaharu Imagama Shizuka Kuno Yoshie Takahashi Masahide Saito Hidehiko Naoe Tomoki |
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Institution: | Department of Hematology, Nagoya University Graduate School of Medicine, Nagoya, Japan. aakihiro@med.nagoya-u.ac.jp |
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Abstract: | We developed a method of clone proliferation promoting fusion genes whose proteins were activated by protein oligomerization through the helix-loop-helix region (PNT domain) of TEL. We inserted a cDNA library downstream of the PNT domain with a retrovirus vector. The resulting retrovirus infected cytokine-dependent 32D cells and cells with cytokine-independent growth were analyzed for the inserted cDNA. We cloned 25 independent fusion genes including seven kinds of partner genes. Six of the seven were a fusion of TEL with protein tyrosine kinase, LYN, HCK, FGR, SYK, FLT3, and TYK2. A serine/threonine kinase, ARAF1, was also found to fuse with TEL. These kinase fusion proteins included kinase domains with proper reading frames. These fusions may be a useful model for clarifying the downstream signal transduction of constitutive active kinase and this expression cloning method may provide a new tool with which to study cell proliferation signalling. |
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Keywords: | Expression cloning Retrovirus vector TEL Tyrosine kinase Serine/threonine kinase |
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