Advanced

Glycerol-3-phosphate transport in Haemophilus influenzae: cloning, sequencing, and transcription analysis of the glpT gene

Song, Xin-Ming; Forsgren, Arne LU and Janson, Håkan LU (1998) In Gene 215(2). p.381-388
Abstract
The presence of a functional glpT gene in Haemophilus influenzae could be questioned, since there is only what appears to be a truncated glpT (HI0686, 143 nt in the 5'-end) available in the H. influenzae Rd genome database (Fleischmann et al. , 1995). For cloning of the glpT gene from H. influenzae type b strain Eagan, an isogenic glpT, rec-1 double mutant and a selective medium for detection of the glpT mutant strains were constructed. The recombinant plasmid carrying glpT was able to complement the isogenic glpT mutant to wild-type levels of G3P uptake and permitted growth on a selective medium with G3P as a major carbon source. The nucleotide sequences of the glpT gene were determined both directly from PCR products and from the cloned... (More)
The presence of a functional glpT gene in Haemophilus influenzae could be questioned, since there is only what appears to be a truncated glpT (HI0686, 143 nt in the 5'-end) available in the H. influenzae Rd genome database (Fleischmann et al. , 1995). For cloning of the glpT gene from H. influenzae type b strain Eagan, an isogenic glpT, rec-1 double mutant and a selective medium for detection of the glpT mutant strains were constructed. The recombinant plasmid carrying glpT was able to complement the isogenic glpT mutant to wild-type levels of G3P uptake and permitted growth on a selective medium with G3P as a major carbon source. The nucleotide sequences of the glpT gene were determined both directly from PCR products and from the cloned DNA insert of strain Eagan. An identical 1440 bp open reading frame with 480 deduced amino acids, highly homologous to other bacterial G3P permeases, was identified. A Northern blot analysis showed that the glpT genes in both Eagan and Rd strains were transcribed on a RNA of approximately 1.4 kb in size. Thus, it is likely that HI0686 sequence originates from a mutated glpT clone in Escherichia coli. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
glp regulon, Fosfomycin resistance, glpT mutant, Selective medium, G3P uptake
in
Gene
volume
215
issue
2
pages
381 - 388
publisher
Elsevier
external identifiers
  • pmid:9714837
  • scopus:0032581499
ISSN
1879-0038
DOI
10.1016/S0378-1119(98)00259-5
language
English
LU publication?
yes
id
204911fc-e268-4765-9b81-ca8b82ddbcbb (old id 1112961)
date added to LUP
2008-07-14 09:23:02
date last changed
2017-02-26 04:13:02
@article{204911fc-e268-4765-9b81-ca8b82ddbcbb,
  abstract     = {The presence of a functional glpT gene in Haemophilus influenzae could be questioned, since there is only what appears to be a truncated glpT (HI0686, 143 nt in the 5'-end) available in the H. influenzae Rd genome database (Fleischmann et al. , 1995). For cloning of the glpT gene from H. influenzae type b strain Eagan, an isogenic glpT, rec-1 double mutant and a selective medium for detection of the glpT mutant strains were constructed. The recombinant plasmid carrying glpT was able to complement the isogenic glpT mutant to wild-type levels of G3P uptake and permitted growth on a selective medium with G3P as a major carbon source. The nucleotide sequences of the glpT gene were determined both directly from PCR products and from the cloned DNA insert of strain Eagan. An identical 1440 bp open reading frame with 480 deduced amino acids, highly homologous to other bacterial G3P permeases, was identified. A Northern blot analysis showed that the glpT genes in both Eagan and Rd strains were transcribed on a RNA of approximately 1.4 kb in size. Thus, it is likely that HI0686 sequence originates from a mutated glpT clone in Escherichia coli.},
  author       = {Song, Xin-Ming and Forsgren, Arne and Janson, Håkan},
  issn         = {1879-0038},
  keyword      = {glp regulon,Fosfomycin resistance,glpT mutant,Selective medium,G3P uptake},
  language     = {eng},
  number       = {2},
  pages        = {381--388},
  publisher    = {Elsevier},
  series       = {Gene},
  title        = {Glycerol-3-phosphate transport in Haemophilus influenzae: cloning, sequencing, and transcription analysis of the glpT gene},
  url          = {http://dx.doi.org/10.1016/S0378-1119(98)00259-5},
  volume       = {215},
  year         = {1998},
}