Molecular cloning and characterization of human TRB2 gene promoter region
Date Issued
2005
Date
2005
Author(s)
Chen, Han-Jou
DOI
en-US
Abstract
In mammals, the development of hematopoietic system depends on cytokines. These cytokines not only mediate cell proliferation and differentiation, but also enhance the survival of these cells by the suppression of apoptotic pathways. Once the cytokines are deprived, cells undergo apoptosis which has been proved to require de novo RNA and protein synthesis, suggesting that new genes are required to proceed the apoptotic signaling pathway. Our goal is to identify the molecules participating in cytokine withdrawal-induced apoptosis of hematopoietic cells. To achieve this goal, our laboratory used microarray to discover the Drosophila tribbles homolog 2 (TRB2) gene that is up-regulated during cytokine withdraw in the cytokine-dependent TF-1 cell line. Further studies using over-expression and RNA interferences technique strongly suggest that TRB2 indeed participating in cytokine withdrawal-induced apoptosis
Since it is little known about the regulation of cytokine withdrawal-induced genes, the aim of this study is to investigate the regulation of TRB2. In the beginning of this thesis, the human tissue distribution of TRB2 were determined. The accumulated TRB2 RNA level was found to achieve the highest level 24 h after cytokine withdraw. Primer extension and 5’ RACE (rapid amplification of cDNA ends) were then performed to identify the transcription start site of human TRB2. We also performed reporter assay to find that the 2-kb fragment upstream of the translation start site may contain the basal promoter activity and cytokine withdrawal-responsive element. Finally, the mouse genomic library was screened for targeting vector construction. The TRB2 knockout mice are intended to be generated for understanding the physiological functions of TRB2.
Subjects
啟動子
cytokine withdraw
TRB2
tribbles
apoptosis
hemopoietic cells
Type
other
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