Construction of a gene bank of Rhodopseudomonas capsulata using a broad host range DNA cloning system |
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Authors: | Gabriele Klug Gerhart Drews |
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Institution: | (1) Institut für Biologie 2, Mikrobiologie, Albert-Ludwigs-Universität, Schaenzlestr. 1, D-7800 Freiburg, Federal Republic of Germany |
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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.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
Dedicated to Hans-Günter Schlegel on occasion of his 60th birthday |
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Keywords: | Gene bank Plasmids pRK290 pRK2013 Rhodopseudomonas capsulata Reconstitution Phototroph negative mutants Absorption spectra Light harvesting complexes |
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