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Lee Y , Emerson CP , Kim MH
A rapid and simple method for the isolation of mutant variants regulating tissue-specific expression of the TnI gene through drug selection
Applied Biochemistry and Biotechnology. 1995 Dec;55(3) :231-240
AbstractTnINEO 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 genomic rearrangement. This result presented the possibility of isolating factor(s) that activate the muscle-specific TnI promotor simply by screening drug-resistant cells having appropriate mutations.
NotesTimes Cited: 0 English Article TK907 APPL BIOCHEM BIOTECH