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

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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
Author(s)
Young Won Lee; C P Emerson; Myung Hee Kim
Bibliographic Citation
Applied Biochemistry and Biotechnology, vol. 55, no. 3, pp. 231-240
Publication Year
1995
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.
Keyword
drug selectionfast troponin I genegenomic rearrangementmutantmyoblaststage-specific expression
ISSN
0273-2289
Publisher
Springer
DOI
http://dx.doi.org/10.1007/BF02786862
Type
Article
Appears in Collections:
1. Journal Articles > Journal Articles
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