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Rapid Analysis for the Isolation of Novel Genes Encoding Putative Effectors to the Position-specific Regulatory Element of Murine Hoxa-7

 Myungsun Cho  ;  Chuog Shin  ;  Wongi Min  ;  Myoung Hee Kim 
 MOLECULES AND CELLS, Vol.7(2) : 220-225, 1997 
Journal Title
Issue Date
Animals ; DNA, Complementary/genetics ; Embryonic and Fetal Development/genetics ; Gene Expression Regulation, Developmental ; Genes, Homeobox* ; Genes, Reporter ; Homeodomain Proteins/genetics* ; Lac Operon ; Mice ; Molecular Sequence Data ; Promoter Regions, Genetic ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Saccharomyces cerevisiae/genetics
Hox genes are known to play a critical role in pattern formation during vertebrate development by being expressed at the specific time and in the specific position along the antero-posterior body axis. In order to understand the regulatory mechanism for the position-specific expression of murine Hoxa-7, yeast one-hybrid system was applied. DNA fragment conferring a position specificity to the Hoxa-7 gene was placed just upstream from the yeast CYC1 promoter and lacZ gene in a reporter. Selection of LacZ positive clones after cotransformation of the reporter and mouse embryonic cDNA library as an effector, which was designed to be expressed as fusion proteins to the GAlA activation domain, allowed us to isolate putative factors interacting with the position-specific regulatory element of murine Hoxa-7. A total of 28 positive clones were screened from 5 x 105 yeast transformants. About 70% of the clones turned out to be novel and most of the candidate clones selected in this study showed a temporally restricted expression pattern during embryonic development, suggesting that this method could provide an efficient way for isolating novel genes whose expressions are temporally regulated during embryogenesis.
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1. College of Medicine (의과대학) > Dept. of Anatomy (해부학교실) > 1. Journal Papers
Yonsei Authors
Kim, Myoung Hee(김명희) ORCID logo https://orcid.org/0000-0001-5652-1452
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