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

被引:50
作者
Paoli, GC
Morgan, NS
Tabita, FR
Shively, JM
机构
[1] OHIO STATE UNIV,DEPT MICROBIOL,COLUMBUS,OH 43210
[2] CLEMSON UNIV,DEPT BIOL SCI,CLEMSON,SC 29634
关键词
ccb genes; RubisCO; form I; form II; Rhodobacter capsulatus; Calvin cycle;
D O I
10.1007/s002030050281
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
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.
引用
收藏
页码:396 / 405
页数:10
相关论文
共 52 条
[1]  
Ausubel FM., 1995, MOL REPROD DEV, V3rd edn, DOI DOI 10.1002/MRD.1080010210
[2]   A RAPID, SENSITIVE METHOD FOR DETECTION OF ALKALINE-PHOSPHATASE CONJUGATED ANTI-ANTIBODY ON WESTERN BLOTS [J].
BLAKE, MS ;
JOHNSTON, KH ;
RUSSELLJONES, GJ ;
GOTSCHLICH, EC .
ANALYTICAL BIOCHEMISTRY, 1984, 136 (01) :175-179
[3]   A COMPLEMENTATION ANALYSIS OF RESTRICTION AND MODIFICATION OF DNA IN ESCHERICHIA COLI [J].
BOYER, HW ;
ROULLAND.D .
JOURNAL OF MOLECULAR BIOLOGY, 1969, 41 (03) :459-&
[4]  
CHEN JH, 1991, J BIOL CHEM, V266, P20447
[5]   COMPLEMENTATION ANALYSIS AND REGULATION OF CO2 FIXATION GENE-EXPRESSION IN A RIBULOSE 1,5-BISPHOSPHATE CARBOXYLASE-OXYGENASE DELETION STRAIN OF RHODOSPIRILLUM-RUBRUM [J].
FALCONE, DL ;
TABITA, FR .
JOURNAL OF BACTERIOLOGY, 1993, 175 (16) :5066-5077
[6]   EXPRESSION OF ENDOGENOUS AND FOREIGN RIBULOSE 1,5-BISPHOSPHATE CARBOXYLASE-OXYGENASE (RUBISCO) GENES IN A RUBISCO DELETION MUTANT OF RHODOBACTER-SPHAEROIDES [J].
FALCONE, DL ;
TABITA, FR .
JOURNAL OF BACTERIOLOGY, 1991, 173 (06) :2099-2108
[7]   A TECHNIQUE FOR RADIOLABELING DNA RESTRICTION ENDONUCLEASE FRAGMENTS TO HIGH SPECIFIC ACTIVITY [J].
FEINBERG, AP ;
VOGELSTEIN, B .
ANALYTICAL BIOCHEMISTRY, 1983, 132 (01) :6-13
[8]   REPLICATION OF AN ORIGIN-CONTAINING DERIVATIVE OF PLASMID RK2 DEPENDENT ON A PLASMID FUNCTION PROVIDED IN TRANS [J].
FIGURSKI, DH ;
HELINSKI, DR .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1979, 76 (04) :1648-1652
[9]   REFINEMENT OF THE HIGH-RESOLUTION PHYSICAL AND GENETIC-MAP OF RHODOBACTER-CAPSULATUS AND GENOME SURVEYS USING BLOTS OF THE COSMID ENCYCLOPEDIA [J].
FONSTEIN, M ;
KOSHY, EG ;
NIKOLSKAYA, T ;
MOURACHOV, P ;
HASELKORN, R .
EMBO JOURNAL, 1995, 14 (08) :1827-1841
[10]   ISOLATION OF THE RHODOPSEUDOMONAS-SPHAEROIDES FORM I RIBULOSE 1,5-BIPHOSPHATE CARBOXYLASE OXYGENASE LARGE AND SMALL SUBUNIT GENES AND EXPRESSION OF THE ACTIVE HEXADECAMERIC ENZYME IN ESCHERICHIA-COLI [J].
GIBSON, JL ;
TABITA, FR .
GENE, 1986, 44 (2-3) :271-278