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Construction of a physical map of the 45 kb photosynthetic gene cluster of Rhodobacter sphaeroides
Authors:S A Coomber  C N Hunter
Institution:(1) Department of Pure and Applied Biology, Imperial College, Prince Consort Road, SW7 2BB London, UK;(2) Present address: Biochemistry Section, Division of Molecular Biology and Biotechnology, Sheffield University, Western Bank, S10 2TN Sheffield, UK
Abstract:1) A number of overlapping clones have been isolated from a Rhodobacter sphaeroides gene bank. Following conjugative gene transfer from Escherichia coli these clones restore a wild type phenotype to several mutants unable to synthesise bacteriochlorophyll. 2) The insert DNA was analysed by restriction mapping and together the clones form the basis of the first restriction map of the 45 kb photosynthetic gene cluster of Rb. sphaeroides. 3) This cluster is bounded on one side by puh A encoding the reaction centre H polypeptide and on the other by the puf operon encoding reaction centre L and M apoproteins and light harvesting LH1 and polypeptides. 4) DNA fragments from the 45 kb cluster were used to probe genomic DNA from other photosynthetic bacteria. Using heterologous hybridisation conditions, a significant degree of homology is shown between Rb. sphaeroides and these other bacteria, suggesting close evolutionary links with Rb. capsulatus in particular.
Keywords:Photosynthesis  Photosynthetic bacteria  Evolution  Bacteriochlorophyll  Reaction centres  Light harvesting
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