Development of a gene cloning system for the hydrogen-producing marine photosynthetic bacterium Rhodopseudomonas sp.

Tadashi Matsunaga, N. Matsunaga, K. Tsubaki, T. Tanaka

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Seventy-six strains of marine photosynthetic bacteria were analyzed by agarose gel electrophoresis for plasmid DNA content. Among these strains, 12 carried two to four different plasmids with sizes ranging from 3.1 to 11.0 megadaltons. The marine photosynthetic bacterium Rhodopseudomonas sp. NKPB002106 had two plasmids, pRD06S and pRD06L. The smaller plasmid, pRD06S, had a molecular weight of 3.8 megadaltons and was cut at a single site by restriction endonucleases SalI, SmaI, PstI, XhoI, and BglII. Moreover, the marine photosynthetic bacterium Rhodopseudomonas sp. NKPB002106 containing plasmid pRD06 had a satisfactory growth rate (doubling time, 7.5 h), a hydrogen-producing rate of 0.96 μmol/mg (dry weight) of cells per h, and nitrogen fixation capability. Plasmid pRD06S, however, had neither drug resistance nor heavy-metal resistance, and its copy number was less than 10. Therefore, a recombinant, plasmid consisting of pRD06S and Escherichia coli cloning vector pUC13 was constructed and cloned in E. coli. The recombinant plasmid was transformed into Rhodopseudomonas sp. NKPB002106. As a result, Rhodopseudomonas sp. NKPB002106 developed ampicillin resistance. Thus, a shuttle vector for gene transfer was constructed for marine photosynthetic bacteria.

Original languageEnglish
Pages (from-to)460-463
Number of pages4
JournalJournal of Bacteriology
Volume168
Issue number1
Publication statusPublished - 1986
Externally publishedYes

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Rhodopseudomonas
Organism Cloning
Hydrogen
Plasmids
Bacteria
Genes
Genetic Vectors
Escherichia coli
Ampicillin Resistance
Nitrogen Fixation
Agar Gel Electrophoresis
DNA Restriction Enzymes
Heavy Metals
Drug Resistance
Molecular Weight
Weights and Measures

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Development of a gene cloning system for the hydrogen-producing marine photosynthetic bacterium Rhodopseudomonas sp. / Matsunaga, Tadashi; Matsunaga, N.; Tsubaki, K.; Tanaka, T.

In: Journal of Bacteriology, Vol. 168, No. 1, 1986, p. 460-463.

Research output: Contribution to journalArticle

Matsunaga, Tadashi ; Matsunaga, N. ; Tsubaki, K. ; Tanaka, T. / Development of a gene cloning system for the hydrogen-producing marine photosynthetic bacterium Rhodopseudomonas sp. In: Journal of Bacteriology. 1986 ; Vol. 168, No. 1. pp. 460-463.
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