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The Application of Yeast Three-Hybrid Technology in the Mexican Axolotl (Ambystoma mexicanum) System.

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Date Issued:
2007
Summary:
The Mexican axolotl, Ambystoma mexicanum, possesses a naturally-occurring lethal mutation, designated gene "c", for cardiac non-function. Hearts form but fail to beat, lack organized myofibrils, and are deficient in tropomyosin. Treatment with a noncoding RNA MIR (Myofibril-Inducing RNA) rescues this defect in organ culture. Rescued mutant hearts have restored tropomyosin, form organized myofibrils, and beat vigorously. Studies to elucidate the mechanism of MIR heart rescue are underway. Current evidence suggests that MIR acts by binding with at least two proteins. The yeast three-hybrid system is being used to screen an axolotl eDNA library for these two proteins and other possible MIR-binding candidates. This is a method utilizing two hybrid proteins and a hybrid RNA. An interaction between these three components will activate the expression of reporter genes, whose activity is assayed through phenotypical and biochemical methods. In this study, the protocol for yeast three-hybrid technology is being established for analyzing the MIR in the Mexican axolotl, cardiac mutant animal model.
Title: The Application of Yeast Three-Hybrid Technology in the Mexican Axolotl (Ambystoma mexicanum) System.
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Name(s): Maier, Jennifer A., author
Lemanski, Larry F., Thesis advisor
Florida Atlantic University, Degree grantor
Type of Resource: text
Genre: Electronic Thesis Or Dissertation
Date Created: 2007
Date Issued: 2007
Publisher: Florida Atlantic University
Place of Publication: Boca Raton, Fla.
Physical Form: application/pdf
Extent: 50 p.
Language(s): English
Summary: The Mexican axolotl, Ambystoma mexicanum, possesses a naturally-occurring lethal mutation, designated gene "c", for cardiac non-function. Hearts form but fail to beat, lack organized myofibrils, and are deficient in tropomyosin. Treatment with a noncoding RNA MIR (Myofibril-Inducing RNA) rescues this defect in organ culture. Rescued mutant hearts have restored tropomyosin, form organized myofibrils, and beat vigorously. Studies to elucidate the mechanism of MIR heart rescue are underway. Current evidence suggests that MIR acts by binding with at least two proteins. The yeast three-hybrid system is being used to screen an axolotl eDNA library for these two proteins and other possible MIR-binding candidates. This is a method utilizing two hybrid proteins and a hybrid RNA. An interaction between these three components will activate the expression of reporter genes, whose activity is assayed through phenotypical and biochemical methods. In this study, the protocol for yeast three-hybrid technology is being established for analyzing the MIR in the Mexican axolotl, cardiac mutant animal model.
Identifier: FA00000793 (IID)
Degree granted: Thesis (M.S.)--Florida Atlantic University, 2007.
Subject(s): Axolotls--Development
Heart--Hypertrophy
Genetic translation
Molecular genetics--Research
Held by: Florida Atlantic University Libraries
Sublocation: Digital Library
Persistent Link to This Record: http://purl.flvc.org/fau/fd/FA00000793
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Host Institution: FAU
Is Part of Series: Florida Atlantic University Digital Library Collections.