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Construction of a gene bank of Rhodopseudomonas capsulata using a broad host range DNA cloning system

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Abstract

A gene bank of the phototrophic bacterium Rhodopseudomonas capsulata was constructed using the binary plasmid system pRK290/pRK2013. Fragments of about 20 kb of chromosomal DNA of R. capsulata strain 37b4 were inserted into the cloning vector pRK290. The hybrid plasmids of the gene bank, maintained in Escherichia coli HB101 were transferred by conjugation to R. capsulata strains defective in the photosynthetic apparatus with frequencies of 5×10-4 to 5×10-2. Phototrophically growing transconjugants occurred with frequencies of 5×10-7 to 5×10-6. Recombination between the hybrid plasmids and the R. capsulata chromosome was shown. The hybrid plasmid pRCF1002, carrying a 25 kb insert of R. capsulata wild type DNA, was isolated from one E. coli clone of the gene bank. It reconstituted some bacteriochlorophyll- and photosynthetic negative mutants to phototrophic growth.

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Abbreviations

Bchl:

Bacteriochlorophyll

RC:

reaction center

LH:

light-harvesting complex

Crt:

carotenoid

pho:

phototrophic growth

P:

Bchl precursor excreted, the number behind P indicates the maximum of absorption in ether (nm)

SDS:

sodium dodecyl sulfate

Tc:

tetracycline

Km:

kanamycin

Gm:

gentamicin

r:

resistant

kb:

kilo base pairs

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Authors and Affiliations

  1. Institut für Biologie 2, Mikrobiologie, Albert-Ludwigs-Universität, Schaenzlestr. 1, D-7800, Freiburg, Federal Republic of Germany

    Gabriele Klug & Gerhart Drews

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  1. Gabriele Klug
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  2. Gerhart Drews
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Dedicated to Hans-Günter Schlegel on occasion of his 60th birthday

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Klug, G., Drews, G. Construction of a gene bank of Rhodopseudomonas capsulata using a broad host range DNA cloning system. Arch. Microbiol. 139, 319–325 (1984). https://doi.org/10.1007/BF00408373

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  • DOI: https://doi.org/10.1007/BF00408373

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