Abstract：
:A yeast transcriptional activator, Gcn4p, induces the expression of genes that are involved in amino acid and purine biosynthetic pathways under amino acid starvation. Gcn4p has an acidic activation domain in the central region and a bZIP domain in the C-terminus that is divided into the DNA-binding motif and dimerization leucine zipper motif. In order to identify amino acids in the DNA-binding motif of Gcn4p which are involved in transcriptional activation, we constructed mutant libraries in the DNA-binding motif through an innovative application of random mutagenesis. Mutant library made by oligonucleotides which were mutated randomly using the Poisson distribution showed that the actual mutation frequency was in good agreement with expected values. This method could save the time and effort to create a mutant library with a predictable mutation frequency. Based on the studies using the mutant libraries constructed by the new method, the specific residues of the DNA-binding domain in Gcn4p appear to be involved in the transcriptional activities on a conserved binding site.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Seong KM,Park H,Kim SJ,Ha HN,Lee JY,Kim Jdoi
10.1016/j.mimet.2007.02.010subject
Has Abstractpub_date
2007-06-01 00:00:00pages
442-50issue
3eissn
0167-7012issn
1872-8359pii
S0167-7012(07)00065-6journal_volume
69pub_type
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