PCR based gene engineering of the Vibrio harveyi lux operon and the Escherichia coli trp operon provides for biochemically functional native and fused gene products

1991 ◽  
Vol 226-226 (1-2) ◽  
pp. 41-48 ◽  
Author(s):  
Philip J. Hill ◽  
Simon Swift ◽  
Gordon S. A. B. Stewart
Keyword(s):  
Escherichia Coli ◽  
Vibrio Harveyi ◽  
Gene Products ◽  
Gene Engineering ◽  
Lux Operon ◽  
Fused Gene ◽  
Trp Operon

scholarly journals Physical and genetic characterization of the melibiose operon and identification of the gene products in Escherichia coli.

1984 ◽  
Vol 259 (3) ◽  
pp. 1807-1812 ◽  
Author(s):  
M Hanatani ◽  
H Yazyu ◽  
S Shiota-Niiya ◽  
Y Moriyama ◽  
H Kanazawa ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Genetic Characterization ◽  
Gene Products

scholarly journals Identification of the Gene Encoding Sulfopyruvate Decarboxylase, an Enzyme Involved in Biosynthesis of Coenzyme M

2000 ◽  
Vol 182 (17) ◽  
pp. 4862-4867 ◽  
Author(s):  
Marion Graupner ◽  
Huimin Xu ◽  
Robert H. White
Keyword(s):  
Escherichia Coli ◽  
Second Step ◽  
Gene Products ◽  
Methanococcus Jannaschii ◽  
Coenzyme M ◽  
Gene Encoding ◽  
Fourth Step

ABSTRACT The products of two adjacent genes in the chromosome ofMethanococcus jannaschii are similar to the amino and carboxyl halves of phosphonopyruvate decarboxylase, the enzyme that catalyzes the second step of fosfomycin biosynthesis inStreptomyces wedmorensis. These two M. jannaschii genes were recombinantly expressed inEscherichia coli, and their gene products were tested for the ability to catalyze the decarboxylation of a series of α-ketoacids. Both subunits are required to form an α6β6 dodecamer that specifically catalyzes the decarboxylation of sulfopyruvic acid to sulfoacetaldehyde. This transformation is the fourth step in the biosynthesis of coenzyme M, a crucial cofactor in methanogenesis and aliphatic alkene metabolism. The M. jannaschiisulfopyruvate decarboxylase was found to be inactivated by oxygen and reactivated by reduction with dithionite. The two subunits, designated ComD and ComE, comprise the first enzyme for the biosynthesis of coenzyme M to be described.

Evidence for two recA genes mediating DNA repair in Bacillus megaterium

Microbiology ◽  
2005 ◽  
Vol 151 (3) ◽  
pp. 775-787 ◽  
Author(s):  
Hannes Nahrstedt ◽  
Christine Schröder ◽  
Friedhelm Meinhardt
Keyword(s):  
Escherichia Coli ◽  
Dna Repair ◽  
Mitomycin C ◽  
Bacillus Megaterium ◽  
Functional Complementation ◽  
Homologous Gene ◽  
Gene Products ◽  
Null Mutant ◽  
Promoter Regions ◽  
Increased Sensitivity

Isolation and subsequent knockout of a recA-homologous gene in Bacillus megaterium DSM 319 resulted in a mutant displaying increased sensitivity to mitomycin C. However, this mutant did not exhibit UV hypersensitivity, a finding which eventually led to identification of a second functional recA gene. Evidence for recA duplicates was also obtained for two other B. megaterium strains. In agreement with potential DinR boxes located within their promoter regions, expression of both genes (recA1 and recA2) was found to be damage-inducible. Transcription from the recA2 promoter was significantly higher than that of recA1. Since a recA2 knockout could not be achieved, functional complementation studies were performed in Escherichia coli. Heterologous expression in a RecA null mutant resulted in increased survival after UV irradiation and mitomycin C treatment, proving both recA gene products to be functional in DNA repair. Thus, there is evidence for an SOS-like pathway in B. megaterium that differs from that of Bacillus subtilis.

scholarly journals Genetic Organization of the Vibrio harveyi dnaA Gene Region and Analysis of the Function of the V. harveyi DnaA Protein in Escherichia coli

2002 ◽  
Vol 184 (9) ◽  
pp. 2533-2538 ◽  
Author(s):  
Dvora Berenstein ◽  
Kirsten Olesen ◽  
Christian Speck ◽  
Ole Skovgaard
Keyword(s):  
Escherichia Coli ◽  
Promoter Region ◽  
Vibrio Harveyi ◽  
Gene Region ◽  
Chromosomal Dna ◽  
Dnaa Gene ◽  
E Coli ◽  
Dnaa Protein ◽  
Dam Methylase ◽  
And Function

ABSTRACT The Vibrionaceae family is distantly related to Enterobacteriaceae within the group of bacteria possessing the Dam methylase system. We have cloned, sequenced, and analyzed the dnaA gene region of Vibrio harveyi and found that although the organization of the V. harveyi dnaA region differs from that of Escherichia coli, the expression of both genes is autoregulated and ATP-DnaA binds cooperatively to ATP-DnaA boxes in the dnaA promoter region. The DnaA proteins of V. harveyi and E. coli are interchangeable and function nearly identically in controlling dnaA transcription and the initiation of chromosomal DNA replication despite the evolutionary distance between these bacteria.

scholarly journals POLARITY IN SEGMENTS OF THE ESCHERICHIA COLI trp OPERON WITH DELETED INTRAOPERONIC TRANSLATIONAL INITIATION SIGNALS

Genetics ◽  
1975 ◽  
Vol 80 (4) ◽  
pp. 651-666
Author(s):  
Yasunobu Kano ◽  
Fumio Imamoto
Keyword(s):  
Escherichia Coli ◽  
Translation Initiation ◽  
Nonsense Mutation ◽  
Messenger Rna ◽  
Distal Part ◽  
High Efficiency ◽  
Translational Initiation ◽  
Transcriptional Termination ◽  
Trp Operon

ABSTRACT The effect of deletion of the operator-distal genes of the trp operon, including the trpE-trpD intercistronic punctuation point, on the degree of transcriptional polarity (in this case the effect of a nonsense mutation on the level of mRNA from the distal part of the very gene where the mutation is located) was investigated. Double mutants which contain a nonsense mutation and a deletion in trpE were constructed, and the degree of transcriptional polarity was estimated by the decrease in messenger RNA for the operator-distal trpE beyond the nonsense mutation, as well as by the production of truncated messenger RNA for the region of trpE proximal to the nonsense mutation. The content of mRNA of operator-distal trpE and the size of the mRNA of operator-proximal trpE of the double mutants show that transcriptional polarity is not relaxed as a function of distance of the nonsense mutation from the operator-distal end of the trpE segment (at which the subsequent high efficiency translational initiation signal has been deleted). These findings are consistent with the conclusion that the degree of polarity depends on the distance of the nonsense mutation fro mthe subsequent translation initiation signal, but not on its distance from the operator-distal end, including possible translational or transcriptional termination signals

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