A rapid and simple method for the isolation of mutant variants regulating tissue-specific expression of the TnI gene through drug selection
- Korea Research Institute of Bioscience and Biotechnology, Taejon (Korea, Republic of)
- Fox Chase Cancer Center, Philadelphia, PA (United States)
TnINEO fusion gene was constructed by fusing 3.4-kbp of quail TnI genomic DNA sequences spanning the promoter to exon 5 and a neo gene in frame. A myoblast cell line was established after transfection of pTnINEO. Since this cell line was passaged several times, a high frequency of neomycin (G418) sensitivity conversion was detected. Two drug-resistant variants were analyzed through genomic Southern blot and S1 nuclease protection assay. One variant has a mutation(s) in the regulatory element that activated the dormant TnI promoter-enhancer in myoblast, and the other has shown the geonomic rearrangement. This result presented the possibility of isolating factor(s) that activate the muscle-specific TnI promoter simply by screening drug-resistant cells having appropriate mutations. 12 refs., 4 fig.
- Sponsoring Organization:
- USDOE
- OSTI ID:
- 254425
- Journal Information:
- Applied Biochemistry and Biotechnology, Vol. 55, Issue 3; Other Information: PBD: Dec 1995
- Country of Publication:
- United States
- Language:
- English
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