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Cloning, sequencing and expression in Escherichia coli of a Streptomyces aureofaciens gene encoding
J Kormanec1, A Lempelová, M Farkasovský
1Institute of Molecular Biology, Slovak Academy of Sciences, Bratislava, Slovak Republic.
Abstract:
The structural gene (gap) encoding glyceraldehyde-3-phosphate dehydrogenase (GAPDH; EC 1.2.1.12) from Streptomyces aureofaciens (Sa) has been cloned and sequenced. The predicted gap product consists of 332 amino acids (aa) (35,312 Da), and has considerable homology (up to 52% aa identity) with other bacterial and eukaryotic gap genes. Sequence analysis of the regions flanking gap revealed two incomplete open reading frames encoding proteins similar to the AraC family of bacterial transcriptional regulators and delta (5)-3-ketosteroid isomerase. The Sa gap gene was expressed at a high level in Escherichia coli (Ec). Transformation of the Ec strain resulted in an up to eightfold increase in specific GAPDH activity.