Cloning of an endoglucanase gene fromPseudomonas fluorescens var.cellulosa intoEscherichia coli andPseudomonas fluorescens |
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Authors: | Lejeune André Colson Charles Eveleigh Douglas E |
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Institution: | (1) Department of Biochemistry and Microbiology, Cook College, Rutgers University, 08903 New Brunswick, NJ, U.S.A.;(2) Present address: Laboratoire de Génétique microbienne, Université Catholique de Louvain, Place Croix du Sud 4 (bte 3), B-1348 Louvain-la-Neuve, Belgium |
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Abstract: | Summary An endoglucanase chromosomal gene from the cellulolyticPseudomonas fluorescens var.cellulosa (NCIB 10462) was cloned inEscherichia coli. Chromosomal DNA was partially digested with the restriction enzymeEcoRI and ligated into the broad host-range, mobilizable plasmid pSUP104 that had been linearized with the same enzyme. After transformation ofEscherichia coli, and endoglucanase-positive clone was detected in situ by use of the Congo-red assay procedure. The endoglucanase gene on the recombinant plasmid pRUCL 100 was expressed in the non-cellulolyticPseudomonas 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.Abbreviations CMC
carboxymethylcellulose
- EG
endoglucanase
- kb
kilobase pairs
- Mops
4-morpholinepropanesulfonic acid
- Apr-s
resistance-sensitivity to the antibiotic ampicillin
- Cmr-s
resistance-sensitivity to the antibiotic chloramphenicol
- Tcr-s
resistance-sensitivity to the antibiotic tetracycline
- Smr-s
resistance-sensitivity to the antibiotic streptomycin
- Tpr-s
resistance-sensitivity to the antibiotic trimethoprim |
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Keywords: | Cloning Expression Endoglucanase Pseudomonas fluorescens Restriction mapping |
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