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UBC Theses and Dissertations

The identification of a third recombinant DNA plasmid encoding a Cellulomonas fimi cellulase gene in Escherichia coli Wakarchuk, Warren William


A shotgun cloning of Cellulomonas fimi DNA into Escherichia coli yielded 2 types of recombinant plasmids encoding carboxymethylcellulase (CMCase) activity (23). These clones were designated pEC1 and pEC2. A third type of recombinant plasmid containing a different C. fimi cellulase gene has now been isolated and designated pEC3. The new clone was differentiated from E. coli (pEC1) and E. coli (pEC2) by the following characteristics: the size of the C. fimi DNA insert; its restriction digestion pattern and map; the levels of CMCase in cell free extracts; halo formation on CMC-Congo Red plates; and the migration of CMCase activity in non-denaturing polyacrylamide gels. As with E. coli (pEC1) and E. coli (pEC2), CMCase activity was detected in the periplasmic space of E. coli (pEC3).

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