Difference between revisions of "PMID:4577939"

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!align=left |Citation
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'''Kusch, M and Wilson, TH'''  (1973) Defective lactose utilization by a mutant of Escherichia coli energy-uncoupled for lactose transport. The advantages of active transport versus facilitated diffusion.''Biochim. Biophys. Acta'' '''311''':109-22
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'''No author listed'''  (1973) Defective lactose utilization by a mutant of Escherichia coli energy-uncoupled for lactose transport. The advantages of active transport versus facilitated diffusion. ''Biochim. Biophys. Acta'' '''311''':109-22
 
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!align=left |Abstract
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!align=left align='left' bgcolor='#CCCCFF' |Abstract
 
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No abstract in PubMed
 
No abstract in PubMed
 
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!align=left |Links
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[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=4577939 PubMed]
 
[http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=pubmed&dopt=Abstract&list_uids=4577939 PubMed]
  
 
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!align=left |Keywords
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!align=left align='left' bgcolor='#CCCCFF' |Keywords
 
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Biological Transport; Biological Transport, Active; Carbon Isotopes; Cell Division; Densitometry; Diffusion; Disaccharides; Enzyme Induction; Escherichia coli; Galactosidases; Kinetics; Lactose; Mutation; Transduction, Genetic
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Biological Transport; Biological Transport, Active; Carbon Isotopes; Cell Division; Densitometry; Diffusion; Disaccharides; Enzyme Induction; Escherichia coli/enzymology; Escherichia coli/growth & development; Escherichia coli/metabolism; Galactosidases/biosynthesis; Galactosidases/metabolism; Kinetics; Lactose/metabolism; Mutation; Transduction, Genetic
  
 
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==Main Points of the Paper ==
 
==Main Points of the Paper ==
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*Mutants described in this paper were defective in energy-dependent accumulation of thiogalactosides, but showed no loss of carrier-mediated transport
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{{LitSignificance}}
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*Inducible energy-uncoupled mutant (54-41) was unable to grow on lactose minimal medium. Inducer IPTG permitted normal growth in these cells
 
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*In the energy-uncoupled mutant, the rate of induction of β-galactosidase by lactose and by low concentrations of IPTG was mucher slower than the parent
 
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*X71-54 (constitutive mutant) grew poorly on melibiose, due to the inability of this sugar to induce α-galactosidase
 
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*For normal growth of inducible cells on lactose and melibiose, carrier-mediated entry (facilitated diffusion) of substrates is inadequate; the membrane carriers must also be capable of accumulating the sugars
 
  
 
== Materials and Methods Used ==
 
== Materials and Methods Used ==
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*Parent Strain: X71 (''i<sup>-</sup>z<sup>+</sup>y<sup>+</sup>a<sup>-</sup>, proC<sup>-</sup>, try<sup>-</sup>, B<sub>1</sub><sup>-</sup>'')
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{{LitMaterials}}
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*Additional Strain: X71-54 (''i<sup>-</sup>z<sup>+</sup>y<sup>UN</sup>a<sup>-</sup>'')- energy uncoupled mutant
 
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*Additional Strain: X71-15 (''i<sup>+</sup>z<sup>+</sup>y<sup>+</sup>a<sup>-</sup>'')
 
−
*Additional Strain: X9003 (''i<sup>+</sup>z<sup>-</sup>y<sup>+</sup>a<sup>+</sup>, B<sub>1</sub><sup>-</sup>'')
 
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*Additional Strain: 54-41 (''i<sup>+</sup>z<sup>+</sup>y<sup>UN</sup>a<sup>-</sup>'')
 
−
 
 
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*Additional Strain: 316 (''i<sup>-</sup>z<sup>+</sup>y<sup>-</sup>a<sup>-</sup>'')- ''y<sup>-</sup>'' strain
 
−
 
 
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*β-galactosidase assay
 
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*α-galactosidase assay
 
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*Carrier-mediated transport assay
 
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*thiogalactoside transacetylase activity was determined
 
−
 
 
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*P<sub>1</sub> Transduction
 
  
 
==Phenotype Annotations==
 
==Phenotype Annotations==
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{| border="2" cellpadding="4" cellspacing="0" style="margin: 1em 1em 1em 0; border: 1px #aaa solid; border-collapse: collapse;"  id="B5223847d8f7cc"  class=" tableEdit Phenotype_Table_2"   
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!|Species!!Taxon ID!!Strain!!Gene (if known)!!OMP!!Phenotype!!Details!!Evidence!!Notes
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!|Phenotype of!!Taxon Information!!Genotype Information (if known)!!Condition Information!!OMP ID!!OMP Term Name!!ECO ID!!ECO Term Name!!Notes!!Status
−
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''Escherichia coli''
 
−
|
 
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NCBI:562
 
−
|
 
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X71-54
 
−
|
 
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''lacY<sup>UN</sup>''
 
−
|
 
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*Abolished lactose catabolic process
 
−
*Absent growth on minimal salts medium with lactose
 
−
*Abolished utilization of carbon source (lactose)
 
−
 
 
−
 
 
−
|
 
−
Growth
 
−
|
 
−
inability to grow on lactose in a minimal salts medium
 
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|
 
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Other
 
−
|
 
−
in text, pg. 113
 
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|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
−
|
 
−
316
 
−
|
 
−
''lacY<sup>-</sup>''
 
−
|
 
−
*Decreased rate of alpha-galactosidase expression
 
−
*Abolished induction of alpha-galactosidase expression
 
−
 
 
−
|
 
−
Metabolic Activity
 
−
|
 
−
alpha-gal activity
 
−
|
 
−
Biochemical Assay
 
−
|
 
−
in text- pg. 116
 
−
|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
−
|
 
−
54-41
 
−
|
 
−
''lacY<sup>UN</sup>''
 
−
|
 
−
*Abolished utilization of carbon source (melibiose)
 
−
*Abolished melibiose catabolic process
 
−
*White colonies on MacConkey melibiose indicator plates
 
−
|
 
−
Growth
 
−
|
 
−
inability to ferment 0.2% melibiose
 
−
 
 
−
|
 
−
Plating Assay
 
−
|
 
−
Table 4
 
−
|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
−
|
 
−
54-41
 
−
|
 
−
''lacY<sup>UN</sup>''
 
−
|
 
−
*Decreased intracellular accumulation of small molecule (IPTG)
 
−
*Decreased activity of LacY transporter
 
−
*Increased facilitative diffusion of LacY?
 
−
*Decreased active transport of LacY?
 
−
*Energy-uncoupled transport?
 
−
|
 
−
Other
 
−
|
 
−
*mutant reached a maximum intracellular concentration of 0.1 mM compared to 0.4 mM IPTG for the energy-coupled parent
 
−
*reduced intracellular IPTG accumulation
 
−
|
 
−
Other
 
−
|
 
−
Figure 1- [<sup>14</sup>C]IPTG uptake
 
−
|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
−
|
 
−
54-41
 
−
|
 
−
''lacY<sup>UN</sup>''
 
−
|
 
−
*Decreased induction of beta-galactosidase
 
−
*Decreased rate of expression of beta-galactosidase
 
−
*Decreased lactose catabolic process
 
−
 
 
−
|
 
−
Metabolic Activity
 
−
|
 
−
*reduced rate of beta-galactosidase induction
 
−
|
 
−
Biochemical Assay
 
−
|
 
−
Figure 4- beta-galactosidase activity measured after lactose induction
 
−
 
−
|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
−
|
 
−
X71-54
 
−
|
 
−
''lacY<sup>UN</sup>''
 
−
|
 
−
*Decreased utilization of carbon source (melibiose)
 
−
*Decreased melibiose catabolic process
 
−
*Decreased growth rate on minimal salts medium with melibiose
 
−
|
 
−
Growth
 
−
|
 
−
*reduced growth rate on melibiose in a minimal salts medium
 
−
|
 
−
Plating Assay
 
−
|
 
−
Table 3
 
−
|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
−
|
 
−
X71-54
 
−
|
 
−
''lacY<sup>UN</sup>''
 
−
|
 
−
*Decreased rate of alpha-galactosidase expression
 
−
*Decreased induction of alpha-galactosidase
 
−
|
 
−
Metabolic Activity
 
−
|
 
−
reduced rate of alpha-galactosidase induction
 
−
|
 
−
Biochemical Assay
 
−
|
 
−
Figure 5- alpha-galactosidase activity measured after melibiose induction
 
−
|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
−
|
 
−
54-41
 
−
|
 
−
''lacY<sup>UN</sup>''
 
−
|
 
−
*Decreased affinity for lactose binding
 
−
*Decreased substrate binding
 
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*Decreased transport of small molecules
 
−
|
 
−
Other
 
−
|
 
−
decreased affinity for lactose
 
−
|
 
−
Other
 
−
|
 
−
Table 5- kinetics of transporter for potential inducer molecules
 
−
|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
−
|
 
−
54-41
 
−
|
 
−
''lacY<sup>UN</sup>''
 
−
|
 
−
*Decreased utilization of carbon source (lactose)
 
−
*Decreased lactose catabolic process
 
−
*Light pink colonies on MacConkey lactose indicator plates
 
−
|
 
−
Growth
 
−
|
 
−
reduced ability to ferment 0.2% lactose (fully restored when IPTG is present)
 
−
 
 
−
|
 
−
Plating Assay
 
−
|
 
−
Table 4 
 
−
|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
−
|
 
−
X71-54
 
−
|
 
−
''lacY<sup>UN</sup>''
 
−
|
 
−
Light pink colonies on MacConkey melibiose indicator plates
 
−
|
 
−
Growth
 
−
|
 
−
inability to ferment 0.2% melibiose
 
−
 
 
−
|
 
−
Plating Assay
 
−
|
 
−
Table 4 
 
−
|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
−
|
 
−
X71-54
 
−
|
 
−
''lacY<sup>UN</sup>''
 
−
|
 
−
*Decreased utilization of carbon source (melibiose)
 
−
*Decreased melibiose catabolic activity
 
−
*Light pink colonies on MacConkey melibiose indicator plate
 
−
|
 
−
Growth
 
−
|
 
−
reduced ability to ferment 0.2% melibiose
 
−
 
 
−
|
 
−
Plating Assay
 
−
|
 
−
Table 4 
 
−
|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
−
|
 
−
54-41
 
−
|
 
−
''lacY<sup>UN</sup>''
 
−
|
 
−
*Abolished lactose catabolic process
 
−
*Absent growth on minimal salts medium with lactose
 
−
*Abolished utilization of carbon source (lactose)
 
−
 
 
−
 
 
−
|
 
−
Growth
 
−
|
 
−
no growth on lactose in liquid culture with lactose concentrations b/t 5M and 50mM
 
−
 
 
−
|
 
−
Plating Assay
 
−
|
 
−
in text, pg. 117- addition of 0.5mM IPTG added to lactose medium lead to growth comparable to parent with no constitutive cells
 
−
|-
 
−
|
 
−
''Escherichia coli''
 
−
|
 
−
NCBI:562
 
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|
 
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54-41
 
−
|
 
−
''lacY<sup>UN</sup>''
 
−
|
 
−
*Increased affinity for melibiose
 
−
*Increased small molecule transport
 
−
*Increased melibiose binding
 
−
|
 
−
Other
 
−
|
 
−
 
 
−
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Other
 
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−
Table 5- kinetics of transporter for potential inducer molecules
 
  
 
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==Notes==
 
==Notes==
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==References==
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{{RefHelp}}
 
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<references/>
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[[Category:Publication]]
 
[[Category:Publication]]
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[[Category:To Be Converted]]
 

Latest revision as of 13:16, 1 September 2013

Citation

No author listed (1973) Defective lactose utilization by a mutant of Escherichia coli energy-uncoupled for lactose transport. The advantages of active transport versus facilitated diffusion. Biochim. Biophys. Acta 311:109-22

Abstract

No abstract in PubMed

Links

PubMed

Keywords

Biological Transport; Biological Transport, Active; Carbon Isotopes; Cell Division; Densitometry; Diffusion; Disaccharides; Enzyme Induction; Escherichia coli/enzymology; Escherichia coli/growth & development; Escherichia coli/metabolism; Galactosidases/biosynthesis; Galactosidases/metabolism; Kinetics; Lactose/metabolism; Mutation; Transduction, Genetic

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Main Points of the Paper

Please summarize the main points of the paper.

Materials and Methods Used

Please list the materials and methods used in this paper (strains, plasmids, antibodies, etc).

Phenotype Annotations

See Help:AnnotationTable for details on how to edit this table.
<protect>

Phenotype of Taxon Information Genotype Information (if known) Condition Information OMP ID OMP Term Name ECO ID ECO Term Name Notes Status
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</protect>

Notes

References

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Retrieved from "http://bcbp-ftq4xm2.biobio.tamu.edu/omp/dev/index.php?title=PMID:4577939&oldid=10714"