Characterization of pMa025, a plasmid from the cyanobacterium Microcystis aeruginosa UV025

• Published in: Archives of microbiology Vol. 177; no. 4; pp. 332 - 338
• Main Authors: Wallace, Margaret M, Miller, David W, Raps, Shirley
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer-Verlag 01.04.2002; Berlin Springer
• Subjects: amino acid sequences; Bacteriology; Base Sequence; Biological and medical sciences; carbenicillin; Cyanophyta; DNA, Bacterial - analysis; Escherichia coli; Freshwater; Fundamental and applied biological sciences. Psychology; Genes, Bacterial; Genes, Regulator; genetic vectors; Genetics; Microbiology; Microcystis - genetics; Microcystis - metabolism; Microcystis aeruginosa; Models, Genetic; open reading frames; plasmids; Plasmids - analysis; Replication Origin; start codon; stop codon; tryptophan; Characterization; Plasmid; Microcystis aeruginosa; Nucleotide sequence; Cyanobacteria; Shuttle vector; Bacteria; Gene organization; Replication
• ISSN: 0302-8933; 1432-072X
• DOI: 10.1007/s00203-002-0397-3

The characterization of pMa025, a plasmid isolated from the unicellular, toxin-producing cyanobacterium Microcystis aeruginosa UV025, is described. A recombinant plasmid, pMaL [pMa025-pBluescript II SK(–)] was constructed for mapping, sequencing, and development of shuttle vectors capable of transforming both Escherichia coli and M. aeruginosa. pMa025 is 8,018 bp in length and has a G+C content of 62.3 mol%. Nineteen presumptive ORFs, ORF A – ORF S were identified using ATG or GTG as initiation codons. Fifteen different ORFs, ORF a – ORF o were identified using TGA as a degenerate codon for tryptophan. GTG was the start codon in two-thirds of the putative ORFs when TGA was the termination codon. GTG was the start codon in one-third of the putative ORFs when TGA was used as a codon for tryptophan. The deduced amino acid sequence from ORF j (3,114 bp) was significantly similar to that of a putative plasmid replication protein, RepA, from plasmid pUH24 of Synecho coccus sp. strain PCC7942. M. aeruginosa UV027 and E. coli were transformed to carbenicillin resistance with pMaL-D7, a 6.4-kb hybrid plasmid (3.46 kb pMa025, 2.95 kb pBluescript II) generated from the nested deletion strategy. pMaL-D7 will be used as a shuttle vector.