caa a22 4500 465824439 CHVBK 20180323112145.0 cr unu---uuuuu 170327e19900701xx s 000 0 eng 10.1007/BF00176657 doi (NATIONALLICENCE)springer-10.1007/BF00176657 The d -galactose dehydrogenase gene from Pseudomonas fluorescens: characterization of mutations leading to increased expression in Escherichia coli [Elektronische Daten] [Stefan Fiedler, Peter Buckel] Summary: The gene encoding d-galactose dehydrogenase (gld; E.C. 1.1.1.48) from Pseudomonas fluorescens is poorly expressed when cloned into Escherichia coli. Mutagenesis of the wild-type construct leads to a strong expression of gld in the heterologous host. To investigate the mutational events directing the increase in expression we constructed a gld-lacZ translational fusion which facilitated the isolation of mutants by colony screening. From several independent mutants three point mutations could be identified. They were distinguished by the sequence position of their respective single base-pair substitutions in the 5′-untranslated region of the gld gene and the degree of enhancement of enzyme activity of the gene product. The influence of these mutations on gld gene expression was analysed by S 1 protection analysis which revealed that their effect was at the level of transcription. Springer-Verlag, 1990 Fiedler Stefan Department of Genetics, Boehringer Mannheim GmbH, Nonnenwald 2, D-8122, Penzberg, Federal Republic of Germany aut Buckel Peter Department of Genetics, Boehringer Mannheim GmbH, Nonnenwald 2, D-8122, Penzberg, Federal Republic of Germany aut Applied Microbiology and Biotechnology Springer-Verlag 33/4(1990-07-01), 418-423 0175-7598 33:4<418 1990 33 253 https://doi.org/10.1007/BF00176657 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00176657 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Fiedler Stefan Department of Genetics, Boehringer Mannheim GmbH, Nonnenwald 2, D-8122, Penzberg, Federal Republic of Germany aut NATIONALLICENCE 700 1- Buckel Peter Department of Genetics, Boehringer Mannheim GmbH, Nonnenwald 2, D-8122, Penzberg, Federal Republic of Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer-Verlag 33/4(1990-07-01), 418-423 0175-7598 33:4<418 1990 33 253 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer