Immortalization of embryonic fibroblasts in heat shock transcription factor 1 knockout mouse / 中南大学学报(医学版)
Journal of Central South University(Medical Sciences)
;
(12): 174-177, 2006.
Article
in Chinese
| WPRIM
| ID: wpr-813740
ABSTRACT
OBJECTIVE@#To establish immortalized embryonic fibroblast lines in heat shock transcription factor 1 (HSF1) HSF1-/- and HSF1+/+ mice and to provide experimental models to study the function of HSF1.@*METHODS@#A mammalian expression vector (pSV3neo) containing the SV40 large T antigen was used to transfect the HSF1-/- and HSF1+/+ mouse embryonic fibroblast using Lipofectamine 2000. Colonies were screened by G418 and expanded to immortalized cell lines. PCR was used to detect the integration of the large T antigen with genome in the mouse embryonic fibroblast. Expression of SV40 large T antigen gene in expanded cells was identified by RT-PCR. HSP70 expression was examined by Western blot in the embryonic fibroblast lines.@*RESULTS@#The stable growth and serial propagation were observed in the HSF1-/- and HSF1+/+ cell lines for six months. The mRNA of SV40 T antigen gene expressed in the two cell lines. HSP70 expression could not be induced in the heat-treated HSF1-/- mouse embryo fibroblasts.@*CONCLUSION@#The immortalized cells of HSF1+/+ and HSF1-/- mouse embryo fibroblasts are successfully established.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pharmacology
/
Transcription Factors
/
Cell Line
/
Antigens, Polyomavirus Transforming
/
Mice, Knockout
/
Cell Biology
/
DNA-Binding Proteins
/
Embryo, Mammalian
/
Fibroblasts
/
Heat Shock Transcription Factors
Type of study:
Prognostic study
Limits:
Animals
Language:
Chinese
Journal:
Journal of Central South University(Medical Sciences)
Year:
2006
Type:
Article
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