Cloning of an endoglucanase gene from pseudomonas fluorescens var cellulosa into escherichia coli and pseudomonas fluorescens

Lejeune, A.; Colson, C.; Eveleigh, D.E.

Journal of Industrial Microbiology 1(2): 79-86


ISSN/ISBN: 0169-4146
Accession: 004977089

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An endoglucanase chromosomal gene from the cellulolytic Pseudomonas fluorescens var. cellulosa (NCIB 10462) was cloned in Escherichia coli. Chromosomal DNA was partially digested with the restriction enzyme EcoRI and ligated into the broad host-range, mobilizable plasmid pSUP104 that had been linearized with the same enzyme. After transformation of Escherichia coli, an endoglucanase-positive clone was detected in situ by use of the Congo-red assay procedure. The endoglucanase gene on the recombinant plasmid pRUCL100 was expressed in the non-cellulolytic Pseudomonas fluorescens PF41. The DNA fragment carrying the gene was transferred to the plasmid pBR322, generating plasmids pRUCL150 and pRUCL151, and its restriction map was derived.