A high-copy-number plasmid capable of replication in thermophilic cyanobacteria.

Abstract

A 2.5 kb high-copy-number plasmid, pMA4 in thermophilic cyanobacterium Synechococcus sp. MA4 was isolated and characterized to develop a genetic engineering system for thermophilic cyanobacteria. The copy number of pMA4 was determined to be by densitometry about 350/cell. The pMA4 may be a type of rolling-circle plasmid, because a possible rep gene encoding 34 kD-protein and a consensus sequence of a double-stranded origin nick site of rolling circle plasmids were found in the pMA4 sequence. The pMA4 was electro-introduced into another thermophile, Synechococcus sp. MA19, which is the strongest poly-beta-hydroxybutyrate (PHB) accumulator in photoautotrophic organisms. The pMA4 was incorporated and retained in MA19. These results indicate that pMA4 could be developed as a useful vector for thermophilic cyanobacteria.