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

Author： Miyake Masato Nagai Hiroshi Shirai Makoto Kurane Ryuichiro Asada Yasuo

ISSN： 0273-2289

Source： Applied Biochemistry and Biotechnology, Vol.77, Iss.1-3, 1999-03, pp. : 267-275

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Abstract

A 2.5 kb high-copy-number plasmid, pM A4 in thermophilic cyanobacterium Synechococcus sp. M A4 was isolated and characterized to develop a genetic engineering system for thermophilic cyanobacteria. The copy number of pM A4 was determined to be by densitometry about 350/cell. The pM A4 may be a type of rolling-circle plasmid, because a possible rep gene encoding 34 k D-protein and a consensus sequence of a double-stranded origin nick site of rolling circle plasmids were found in the pM A4 sequence. The pM A4 was electro-introduced into another thermophile, Synechococcus sp. MA 19, which is the strongest poly-β-hydroxybutyrate (PHB) accumulator in photoau totrophic organisms. The pM A4 was incorporated and retained in MA 19. These results indicate that pM A4 could be developed as a useful vector for thermophilic cyanobacteria.