Construction of 293pT2-P210 cell line enables expression of bcr/abl to be regulated by Tet-off inducing-expression-system / 中国实验血液学杂志
Journal of Experimental Hematology
; (6): 224-228, 2007.
Article
在 Zh
| WPRIM
| ID: wpr-230295
Responsible library:
WPRO
ABSTRACT
Chronic myelogenous leukemia (CML) is a clonal myeloproliferative disease of transformed hematopoietic progenitor cells. It is now clear that the chimeric bcr/abl P210(bcr/abl) fusion protein, which is generated by the reciprocal translocation t (9; 22), inhibits apoptosis and increase proliferation. P210(bcr/abl) plays a central role in the pathophysiology of CML. The purpose of this study was to construct a cell line model that bcr/abl expression can be regulated by Tet-off inducing-expression-system. The full-length b3a2 bcr/abl cDNA was subcloned into the pTRE2hyg expression vector to construct the pT2-P210 plasmid. 293 cells were firstly transfected with Tet-off plasmid and the clone that the Tet-off system can work effectively after transfected with pTRE2hyg-LUC was selected by luciferase activity assay. The pT2-P210 plasmid was then transfected into the selected clone and cells were then selected for hygromycin B and G418 resistance. The results showed that individual subclones expressing bcr/abl after withdrawing doxycycline were 293pT2-P210 cell line. In conclusion, selected 293pT2-P210 cells are cells that bcr/abl expression can be regulated by Tet-off inducing-expression-system. They are suitable thoroughly to study the function of bcr/abl fusion gene and its signal regulation mechanism.
全文:
1
索引:
WPRIM
主要主题:
Pathology
/
Physiology
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Translocation, Genetic
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Chromosomes, Human, Pair 9
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Chromosomes, Human, Pair 22
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Cell Line, Transformed
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Molecular Sequence Data
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Tumor Cells, Cultured
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Base Sequence
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Transfection
限制:
Humans
语言:
Zh
期刊:
Journal of Experimental Hematology
年:
2007
类型:
Article