Characterization and expression in @entity526 of an endoglucanase gene of Pseudomonas fluorescens subsp. XXXX .

@entity27928 @entity2975 @entity526
An endoglucanase gene of @entity27928 present on plasmid pRUCL150 and expressed in @entity526 was subcloned in plasmid pBR322. Plasmid pRUCL153 contained the smallest DNA insert (2.9 kb) with endoglucanase activity. The plasmids directed the synthesis of a mostly periplasmic enzyme in @entity526 and the level of enzyme activity was comparable in several strains. Analysis by non-denaturing @entity2975 gel electrophoresis of the endoglucanase produced with various recombinant plasmids showed that it was unique. The endoglucanase gene on plasmid pRUCL153 was localized by physical mapping of independent transposon Tn5 insertions. Hence, its size was estimated to be approx. 1.3 kb. In vivo radioactive labelling of plasmid-encoded proteins using minicells, followed by denaturing @entity2975 gel electrophoresis, allowed us to determine the size of the endoglucanase: Mr 40,000 for the precursor and Mr 38,000 for the mature enzyme. It was demonstrated that no cellulase operon, but a single gene, was cloned. The direction of transcription of the gene was determined by placing it under the control of the promoter of the lactose operon.
