Функциональный анализ rep-области плазмиды pBS267 группы IncP-9, детерминирующей деградацию капролактама

Abstract

The mini-replicon of IncP-9 group plasmid pBS267 (ã-subgroup) controlling caprolactam degradation has been cloned for the first time. It has been determined that minimal replicon containing rep-gene and oriV-site was characterized by the instability of inheritance in bacteria Pseudomonas (in 12 species of Pseudomonas were investigated). Using mutagenesis in vitro the variants of mini-replicon, whose stability of inheritance in bacteria Pseudomonas was increased up to 100 %, have been obtained. The sequences analysis of mutant plasmids has allowed detecting transitions (substitutions of guanine for adenine) in rep-gene and oriV-site region. It has been determined that the mutant variants have been characterized by increase of copy numbers in the cells of bacteria P. putida. The obtained results can serve as a basis for studying the replication mechanism of IncP-9 plasmids, and the obtained constructions can be used for creating vector systems for molecular cloning in a broad range of gram-negative bacteria