Cloning and characterization of chromosomally encoded cephalosporinase gene of Enterobacter cloacae
| Autor: | Sylvain Guerin, Roger Guay, François Paradis |
|---|---|
| Rok vydání: | 1986 |
| Předmět: |
DNA
Bacterial Immunology Enterobacter Biology Molecular cloning Applied Microbiology and Biotechnology Microbiology beta-Lactamases Restriction fragment Plasmid Restriction map Bacterial Proteins Enterobacteriaceae Escherichia coli Genetics Deletion mapping Cloning Molecular Molecular Biology Cephalosporinase General Medicine biology.organism_classification Molecular biology Subcloning Genes Bacterial biology.protein Isoelectric Focusing Hybrid plasmid Enterobacter cloacae Plasmids |
| Zdroj: | Canadian Journal of Microbiology. 32:301-309 |
| ISSN: | 1480-3275 0008-4166 |
| DOI: | 10.1139/m86-061 |
| Popis: | The cephalosporinase gene, cpa, which codes for an inducible class I chromosomal β-lactamase in Enterobacter cloacae was cloned on a fragment of 6.05 kilobase pairs inserted into plasmid pACYC184 and transferred into Escherichia coli HB101 recipient cells. The constructed hybrid plasmid, designated pGGQ101, carried a genomic fragment which retained its parental inducibility characteristics, although its expression level in transformed E. coli cells fell to 40–65% of its initial level in E. cloacae. The localization of the cpa gene on pGGQ101 plasmid was determined by Bal31 exonuclease deletion mapping and further confirmed by subcloning HindIII–AvaI restriction fragment on pMB9 plasmid vector. Labeling with [35S]methionine of pGGQ101 specified proteins in a minicell system showed that six or seven proteins are encoded by the insert. Two proteins with apparent molecular mass of 42 000 and 39 500 daltons, respectively, most probably represent the premature and mature cephalosporinase forms. |
| Databáze: | OpenAIRE |
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