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Applied Biochemistry and Biotechnology

, Volume 55, Issue 3, pp 231–240 | Cite as

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

  • Youngwon Lee
  • Charles P. Emerson
  • Myoung Hee Kim
Original Articles

Abstract

TnINEO fusion gene was constructed by fusing 3.4-kbp of quailTnI genomic DNA sequences spanning the promoter to exon 5 and aneo 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 dormantTnI 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-specificTnI promoter simply by screening drug-resistant cells having appropriate mutations.

Index Entries

Fast troponin I gene stage-specific expression myoblast drug selection mutant genomic rearrangement 

Abbreviations

bp

base pair(s)

kbp

kilobase pair(s)

TnI

troponin I gene

neo

neomycin resistance gene

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Copyright information

© Humana Press Inc 1995

Authors and Affiliations

  • Youngwon Lee
    • 1
  • Charles P. Emerson
    • 2
  • Myoung Hee Kim
    • 1
  1. 1.Genetic Resources CenterKorea Research Institute of Bioscience and BiotechnologyTaejonKorea
  2. 2.Fox Chase Cancer CenterPhiladelphia

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