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PMID: 7721699 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Complete DNA sequence, specific Tn5 insertion map, and gene assignment of the carotenoid biosynthesis pathway of Rhodobacter sphaeroides.

Journal of bacteriology ·Vol. 177 ·No. 8 ·1995-04-00 ·Pages 2064-73

Lang HP, Cogdell RJ, Takaichi S, Hunter CN

Abstract

The carotenoid biosynthesis genes form a cluster within the genome of Rhodobacter sphaeroides, lying in the middle of a larger cluster and 45 kb in length, which contains genes for bacteriochlorophyll biosynthesis and for the reaction center and light-harvesting apoproteins. The positions and approximate limits of the carotenoid genes were determined previously by localized transposon Tn5 mutagenesis and by comparison with the closely related Rhodobacter capsulatus carotenoid gene cluster. In this report, analysis of the DNA and deduced amino acid sequences of the carotenoid genes in R. sphaeroides are presented. Twenty-five Tn5 insertion mutants were used to produce a base-specific Tn5 insertion map of this region, and carotenoid gene assignment was supported by spectroscopic, ultrastructural, and high-pressure liquid chromatography analyses of these mutants. A region in the 3' end of crtD which affects bacteriochlorophyll biosynthesis was discovered, and CrtA was found to possess a proline-rich C-terminal region containing a repeated (Ala-Pro)n motif. CrtF also showed a high degree of sequence conservation with eukaryotic O-methyltransferases. This study provides gene sequences and assignments based upon a comprehensive structural, spectroscopic, and biochemical analysis of a range of carotenoid biosynthetic mutants; in each mutation, the point of Tn5 insertion is determined accurate to 1 bp on the gene cluster.

Related Genes
crtA crtB crtC crtD crtE crtF crtI crtK
MeSH Terms
Amino Acid Sequence Base Sequence Biological Evolution Carotenoids/biosynthesis,genetics Chromosome Mapping Conserved Sequence DNA Transposable Elements DNA, Bacterial/genetics Escherichia coli/genetics Genes, Bacterial Molecular Sequence Data Multigene Family Mutagenesis, Insertional Mutation Rhodobacter sphaeroides/genetics,metabolism
Chemicals
DNA Transposable Elements DNA, Bacterial Carotenoids
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Lang H P
Department of Molecular Biology and Biotechnology, Robert Hill Institute for Photosynthesis and Krebs Institute, University of Sheffield, United Kingdom.
Cogdell R J
Takaichi S
Hunter C N
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.177.8.2064-2073.1995
Published
1995-04-00
Pages
2064-73
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC176850
Subset
IM
Databases
GENBANK
X82458
External Links
PubMed source DOI: 10.1128/jb.177.8.2064-2073.1995 PMC Full Text View journal details
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