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Title 

A rapid and simple method for the isolation of mutant variants regulating tissue-specific expression of the TnI gene through drug selection

Authors 

Young Won LeeC P EmersonMyung Hee Kim

Publisher 

Humana Press

Issue Date 

1995

Citation 

Applied Biochemistry and Biotechnology, vol. 55, no. 3, pp. 231-240

Keywords 

drug selectionfast troponin I genegenomic rearrangementmutantmyoblaststage-specific expression

Abstract 

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 genomic 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.

ISSN 

0273-2289

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2017-04-19


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