Cytoplasmic phosphorylating-domain of the mannitol-specific transport protein of the phosphoenolpyruvate-dependent phosphotransferase system in Escherichia coli: overexpression, purification and functional complementation with the mannitol binding domain

Biochemistry ◽  
1991 ◽  
Vol 30 (39) ◽  
pp. 9478-9485 ◽  
Author(s):  
Rob P. Van Weeghel ◽  
Gert Meyer ◽  
Hendri H. Pas ◽  
Wolfgang Keck ◽  
George T. Robillard
Genetics ◽  
1987 ◽  
Vol 115 (3) ◽  
pp. 419-429
Author(s):  
Maja Kricker ◽  
Barry G Hall

ABSTRACT The cellobiose catabolic system of Escherichia coli K12 is being used to study the role of cryptic genes in microbial evolution. Wild-type E. coli K12 do not utilize the β-glucoside sugars, arbutin, salicin and cellobiose. A Cel+ (cellobiose utilizing) mutant which grows on cellobiose, arbutin, and salicin was isolated previously from wild-type E. coli K12. Biochemical assays indicate that a cel structural gene (celT) specifies a single transport protein that is a β-glucoside specific enzyme of the phosphoenolpyruvate-dependent phosphotransferase system. The transport protein phosphorylates β-glucosides at the expense of phosphoenolpyruvate. A single phosphoglucosidase, specified by celH, hydrolyzes phosphorylated cellobiose, arbutin, and salicin. The genes of the cel system are expressed constitutively in the Cel+ mutant, whereas they are not expressed at a detectable level in the wild-type strain. The transport and hydrolase genes are simultaneously silenced or simultaneously expressed and thus constitute an operon. Cel+ strains which fail to utilize one or more β-glucosides express the transport system at a lower level than do Cel+ strains which grow on all three β-glucosides. Other strains inducibly express a gene which specifies transport of arbutin but not the other β-glucosides. The arbutin transport gene, arbT, maps outside of the cel locus.


1991 ◽  
Vol 266 (31) ◽  
pp. 20922-20927 ◽  
Author(s):  
K. Kashiwagi ◽  
T. Suzuki ◽  
F. Suzuki ◽  
T. Furuchi ◽  
H. Kobayashi ◽  
...  

Genetics ◽  
1998 ◽  
Vol 148 (4) ◽  
pp. 1441-1451
Author(s):  
Johan G de Boer ◽  
Barry W Glickman

Abstract The lacI gene has been used extensively for the recovery and analysis of mutations in bacteria with various DNA repair backgrounds and after exposure to a wide variety of mutagens. This has resulted in a large database of information on mutational mechanisms and specificity of many mutagens, as well as the effect of DNA repair background on mutagenicity. Most importantly, knowledge about the mutational sensitivity of the lacI gene is now available, yielding information about mutable nucleotides. This popularity and available knowledge resulted in the use of the lacI gene in transgenic rodents for the study of mutagenesis in mammals, where it resides in ~40 repeated copies. As the number of sequenced mutations recovered from these animals increases, we are able to analyze the sites at which mutations have been recovered in great detail and to compare the recovered sites between bacteria and transgenic animals. The nucleotides that code for the DNA-binding domain are nearly saturated with base substitutions. Even after determining the sequences of ~10,000 mutations recovered from the animals, however, new sites and new changes are still being recovered. In addition, we compare the nature of deletion mutations between bacteria and animals. Based on the nature of deletions in the animals, we conclude that each deletion occurs in a single copy of the gene.


1991 ◽  
Vol 266 (11) ◽  
pp. 6690-6692
Author(s):  
H H Pas ◽  
G H Meyer ◽  
W H Kruizinga ◽  
K S Tamminga ◽  
R P van Weeghel ◽  
...  

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