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KMID : 0624219980060000024
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Focus on Genetic Science
1998 Volume.6 No. 0 p.24 ~ p.31
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Identification of Trans-activating Factor(s) Binding to the Promoter Region of Human GLUT2 Gene
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Cha, Ji-young
Kim, Ha-il/Kim, Jae-woo/Ahn, Yong-ho
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Abstract
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Previously, we reported that the -44 region is frequently mutated from base A to G in type 2 diabetes mellitus and possibly related to decreased gene expression. Thus, we have designed an experiment to identify transaction factor(s) binding to this region, which may be responsible for the regulation of GLUT2 gene. To this end, we screened human liver cDNA library using yeast one-hybrid system. The cis-element binding the transactivation factors was constructed by synthesizing oligomers repeating the -46/-27 region three times and subcloned into pHisi, pHisis-1, and pLacZi vector. This sequence was incorporated into yeast exploiting homologous recombination phenomenon. In this study, we were able to obtain 63 positive clones and sequenced. Computer analysis of the clones revealed that 6 clones contained possible protein coding sequence regulating human GLUT2 promoter activity. Currently, detailed molecular characterization and biological significance of these proteins are under intensive study.
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KEYWORD
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