Effect of Environmental Factors on the Expression of the Catabolite-Dependent lux-Operon Borne by a Recombinant Plasmid
نویسندگان
چکیده
Expression of the lux-genes cloned in the recombinant plasmid pPHL7 (Ap r Lux + ) in Escherichia coli Z905 cells was studied in various environments, including model aquatic ecosystems. Expression of the lux-genes strongly depended on the nutritional status of the medium. In particular, the cultivation of cells in nutrient-rich medium favored the maintenance of the initial level of expression of the lux-operon, whereas nutrient limitation induced recombinant cell variants with an impaired control of the catabolite-dependent luxoperon. On the other hand, long-term laboratory cultivation of the recombinant strain in nutrient-deficient media or its long-term life in model aquatic ecosystems led to the accumulation of cells with a stringent control on the cloned lux-genes in the bacterial population. The presence of the selective factor (ampicillin) in the medium had no significant effect on the expression of the lux-operon.
منابع مشابه
PCR-MEDIATED CLONING A ND EXPRESSION OF THE GENE FOR THE B-SUBUNIT OF VIBRIO CHOLERAE TOXIN ISOLATED RECENTLY IN IRAN
Knowing the nucleotide sequence of the cholera toxin operon, we designed oligonucleotide primers for its-PCR amplification from local clinical isolates of V. cholerae. The resulting amplification product was cloned in a common pUC18 vector. Subsequently, a part of this operon encoding the cholera toxin Bsubunit (CTB) was reamplified and cloned between the BamH1 and EcoR1 sites of the same ...
متن کاملExpression of Recombinant Protein B Subunit Pili from Vibrio Cholera
Background & Aims: Vibrio cholerae is a gram-negative bacterial pathogen that causes cholera disease. Following ingestion by a host and entry into the upper intestine, V. cholera colonizes and begins to emit enterotoxin. One of the most pathogenic factors of Vibrio cholera is toxin-coregulated pili (TCP). ToxinCoregulated pili is as the primary factor requiered for the colonization and insisten...
متن کاملAn alternative bacterial expression system using Bacillus pumilus SG2 chitinase promoter
Background: Chitin is an abundant natural polysaccharide found in fungi, algae, and exoskeleton of insects. Several bacterial species are capable of utilizing chitin as their carbon source. These bacteria produce chitinases for degradation of chitin into N-acetyl-D-glucosamine. So far, regulation of the chitinase encoding genes has been studied in different bacterial species. Among Bacillus spe...
متن کاملStability of Recombinant Proteins in Escherichia coli: The Effect of Co-Expression of Five Different Chaperone Sets
Chaperones are produced by prokaryotic, yeast and higher eukaryotic cells for various purposes. Over-expression of each chaperone or sets of them affect the production level of a recombinant protein in the cell. On the basis of this hypothesis, five different plasmids with 5 different combinations of 6 chaperones molecule, transformed into Escherichia coli along with human basic Fibroblast Grow...
متن کاملEffects of ackA, pta and poxB inhibition by antisense RNA on acetate excretion and recombinant beta interferon expression in Escherichia coli
Introduction: Escherichia coli (E.coli) is one of the most widely used hosts for the production of recombinant proteins. The main problem in getting high product yields and productivity is the accumulation of acetic acid (acetate) as an unwanted metabolic by-product. In this study, an antisense-based strategy as a metabolic engineering approach was employed to hamper the acetate excretion probl...
متن کامل