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Segregational and structural instability of recombinant plasmid carrying genes for naphthalene degrading pathway
Author(s) -
G. K. Samanta,
Rani Rani,
Alok Jain
Publication year - 1998
Publication title -
letters in applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.698
H-Index - 110
eISSN - 1472-765X
pISSN - 0266-8254
DOI - 10.1046/j.1472-765x.1998.00316.x
Subject(s) - plasmid , pseudomonas putida , recombinant dna , biology , ecori , microbiology and biotechnology , plasmid preparation , molecular cloning , structural gene , t dna binary system , gene , chemistry , genetics , escherichia coli , vector (molecular biology) , peptide sequence , pbr322
The stability of recombinant plasmid carrying genes for naphthalene mineralization was determined. A strain of Pseudomonas putida capable of mineralizing naphthalene (Nap + ) via salicylate (Sal + ) was isolated, and all regulatory and structural genes for the whole pathway were found to be encoded on a 25 kb Eco RI fragment of an approximately 83 kb plasmid present in this strain. The 25 kb Eco RI fragment was cloned into a tetracycline‐resistant (Tc R ) cloning vector pLAFR3 and the recombinant plasmid, pRKJ3 (Nap + , Sal + , Tc R ), thus obtained was transferred into the plasmid‐free strain Pseudomonas putida KT2442 in order to test the stability of the plasmid. Plasmid pRKJ3 was found to be segregationally and/or structurally unstable, depending on the growth conditions. Two types of novel derivative strains having the phenotypes Nap − , Sal + , Tc R and Nap − , Sal − , Tc R with specific deletions of approximately 2 kb and 18 kb, respectively, were obtained.