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KMID : 0903519960390060443
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
1996 Volume.39 No. 6 p.443 ~ p.448
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Regulatory Characterization of xylA Promoter Region in Escherichia coli
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Abstract
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In order to investigate the function of xylA promoter(Pxyl) as regulatory region Pxyl-lacZ fusion gene was constructed by the insertion of xylA promoter to the multiple cloning site of upstream of lacZ gene in a multicopy numbered plasmid pMC1403 containing promoterless lac operon, which was designated pMCX 191, and Pxyl-lacZ fragment from pMCX191 was inserted to low copy numbered plasmid pLG339, designated pLGX191. The expressions of β-galactosidase in these recombinant plasmids containing Pxyl-lacZ fusion gene were induced strongly by the addition of xylose, repressed by the addition of 0.2% glucose in the presence of xylose. The catabolite repressions were derepressed by the addition of 1 mM cAMP as same as native xylA gene. The fragment of xylA promoter was partially deleted from the upstream of xylA promoter by exonuclease Ⅲ to investigate the regulation site of xylA promoter and the degrees of deletion derivatives of xylA promoter were analyzed by the DNA base sequencing. By the investigations of the induction by xylose, repression by glucose and derepression by CAMP on xylose isomerase production, the regulation site of xylA promoter may be located in segment between 165 and -59 by upstream from the initiation site of xylA translation.
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