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PMID: 8588741 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Expression of the cbbLcbbS and cbbM genes and distinct organization of the cbb Calvin cycle structural genes of Rhodobacter capsulatus.

Archives of microbiology ·Vol. 164 ·No. 6 ·1995-12-00 ·Pages 396-405

Paoli GC, Morgan NS, Tabita FR, Shively JM

Abstract

Rhodobacter capsulatus fixes CO2 via the Calvin reductive pentose phosphate pathway and, like some other nonsulfur purple bacteria, is known to synthesize two distinct structural forms of ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO). Cosmid clones that hybridized to form I (cbbLcbbS) and form II (cbbM) RubisCO gene probes were isolated from a genomic library of R. capsulatus strain SB1003. Southern blotting and hybridization analysis with gene-specific probes derived from Rhodobacter sphaeroides revealed that R. capsulatus cbbM is clustered with genes encoding other enzymes of the Calvin cycle, including fructose 1,6/sedoheptulose 1,7-bisphosphatase (cbbF), phosphoribulokinase (cbbP), transketolase (cbbT), glyceraldehyde-3-phosphate dehydrogenase (cbbG), and fructose 1,6-bisphosphate aldolase (cbbA), as well as a gene (cbbR) encoding a divergently transcribed LysR-type regulatory protein. Surprisingly, a cosmid clone containing the R. capsulatus form I RubisCO genes (cbbL and cbbS) failed to hybridize to the other cbb structural gene probes, unlike the situation with the closely related organism R. sphaeroides. The form I and form II RubisCO genes were cloned into pUC-derived vectors and were expressed in Escherichia coli to yield active recombinant enzyme in each case. Complementation of a RubisCO-deletion strain of R. sphaeroides to photosynthetic growth by R. capsulatus cbbLcbbS or cbbM was achieved using the broad host-range vector, pRK415, and R. sphaeroides expression vector pRPS-1.

MeSH Terms
Chromosome Mapping Cloning, Molecular Cosmids Escherichia coli/genetics Gene Expression Genes, Bacterial Genetic Complementation Test Multigene Family Pentose Phosphate Pathway/genetics Rhodobacter capsulatus/genetics,metabolism Ribulose-Bisphosphate Carboxylase/genetics
Chemicals
Ribulose-Bisphosphate Carboxylase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Paoli G C
Department of Microbiology, The Ohio State University, Columbus, OH 43210, USA.
Morgan N S
Tabita F R
Shively J M
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Article Info
Journal
Archives of microbiology
Abbr.
Arch Microbiol
ISSN
0302-8933
Published
1995-12-00
Pages
396-405
Language
English
Region
Germany
NLM ID
0410427
Subset
IM
Grants
NIGMS NIH HHS · GM-45404 · United States
External Links
PubMed source View journal details
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