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KMID : 0903519950380020111
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
1995 Volume.38 No. 2 p.111 ~ p.117
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Nucleotide Sequence and Cloning of sfs4 , One of the Genes Involved in the CRP - Dependent Expression of E . coli mad Genes
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Abstract
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In Escherichia coli, CRP forms a complex with cAMP and acts as a transcriptional regulator of many genes, including sugar metabolism operons. The E coli MK2001, which is introduced the altered crp^(*1), is functional in the expression of lac, ara and man, in the absence of cAMP. However the expression of mat gene is fully activated by the addition of cAMP or cGMP. The object of the study is cloning of the sfs (sugar fermentation stimulation) genes, which was involved in regulation of mat gene expression with the altered crp^(*1) gene, and structural analysis and characterization of the genes at the molecular level. We have cloned 5 different E. coli genes which stimulate the maltose metabolism in a crp^(*1), cya::km (MK2001) background. Newly identified genes were designated as sfs. One of the sfs genes (pPC1), located at the 53.2 min map position on the E coli chromosome, was further analyzed. Expression of the genes, which is involved in maltose metabolism, malQ (amylomaltase), was increased to 5.8-fold in the presence of a plasmid, pAPS, containing the subcloned sfs4 gene. The nucleotide seguence of a common 2,126 by segment of the pPCM1 was determined and two open reading frames (ORF1 and ORF2) were detected. The ORF1 encodes the sfs4 gene and ORF2 encodes a truncated protein. Potential CRP binding site is located in the upstream of the putative promoter in the regulatory region. Expression of the cloned sfs4 gene was positively regulated by the cAMP-CRP complex.
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