Difference between revisions of "PMID:4887874"

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!align=left  |Keywords
 
!align=left  |Keywords
 
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Acyltransferases; Chromosome Mapping; Escherichia coli; Genes; Membrane Transport Proteins; Mutation; Operon
+
Polar Effect; Acyltransferases; Chromosome Mapping; Escherichia coli; Genes; Membrane Transport Proteins; Mutation; Operon; Translation; Suppressor
  
 
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== Materials and Methods Used ==
 
== Materials and Methods Used ==
 
*Parent Strain: NG545 (''trp-8'', Sm<sup>S</sup> Hfr)
 
*Parent Strain: NG545 (''trp-8'', Sm<sup>S</sup> Hfr)
 +
**UAG mutation had not been altered in the suppressor strains isolated
 
*Strain: CA77B (''Δlac, trp-8'', Sm<sup>R</sup> F<sup>-</sup>)
 
*Strain: CA77B (''Δlac, trp-8'', Sm<sup>R</sup> F<sup>-</sup>)
 
*Strain: X103B (''Δlac pro, trp-8'', Sm<sup>R</sup> F<sup>-</sup>)
 
*Strain: X103B (''Δlac pro, trp-8'', Sm<sup>R</sup> F<sup>-</sup>)
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*Strain: ΔLac<sub>W</sub> (''Δlac, his<sub>-</sub>'', Sm<sup>R</sup> F<sup>-</sup>)
 
*Strain: ΔLac<sub>W</sub> (''Δlac, his<sub>-</sub>'', Sm<sup>R</sup> F<sup>-</sup>)
 
*Strain: E29 (''lac<sup>+</sup> '', F'/''lac<sup>+</sup>'', threonine<sup>-</sup>, leucine<sup>-</sup>, B<sub>1</sub><sup>-</sup>, Sm<sup>R</sup>)
 
*Strain: E29 (''lac<sup>+</sup> '', F'/''lac<sup>+</sup>'', threonine<sup>-</sup>, leucine<sup>-</sup>, B<sub>1</sub><sup>-</sup>, Sm<sup>R</sup>)
 
  
 
*Acetylase assay
 
*Acetylase assay
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lacZ  
 
lacZ  
 
|
 
|
able to metabolize melibiose
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*Growth on melibiose medium
 +
*Melibiose catabolic process
 +
 
 
|
 
|
 
Growth
 
Growth
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lacZ  
 
lacZ  
 
|
 
|
 
+
*Suppressor-dependent growth on lactose medium
 +
*Suppressor-dependent lactose catabolic process
 
|
 
|
 
Growth
 
Growth
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lacZ  
 
lacZ  
 
|
 
|
unable to metabolize melibiose
+
*Abolished melibiose catabolic process
 +
*Absent growth on melibiose medium
 
|
 
|
 
Growth
 
Growth
 
|
 
|
growth on melibiose-polar effects
+
unable to metabolize melibiose/ growth on melibiose-polar effects
 
|
 
|
 
Plating Assay
 
Plating Assay
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lacZ  
 
lacZ  
 
|
 
|
reduced beta-galactosidase activity
+
*Decreased gene expression of beta-galactosidase
 +
*Decreased lactose catabolic process
 
|
 
|
 
Metabolic Activity
 
Metabolic Activity
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lacZ  
 
lacZ  
 
|
 
|
reduced acetylase activity
+
*Decreased acetylase activity
 +
*Decreased lactose catabolic process
 
|
 
|
 
Metabolic Activity
 
Metabolic Activity
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NCBI:562
 
NCBI:562
 
|
 
|
NG545-pi(1)<sub>a</sub>B
+
NG545
 
|
 
|
 
lacZ  
 
lacZ  
 
|
 
|
reduced acetylase activity
+
*Suppressor-dependent growth on lactose medium
 +
*Suppressor-dependent lactose catabolic process
 +
|
 +
Growth
 +
|
 +
able to metabolize lactose only in a su<sub>I</sub><sub>+</sub> background
 +
|
 +
Plating Assay
 +
|
 +
Table 2- UAG polar mutation 
 +
|-
 +
|
 +
''Escherichia coli''
 +
|
 +
NCBI:562
 +
|
 +
NG545-pi(1)
 +
|
 +
lacZ
 +
|
 +
*Decreased acetylase activity
 +
*Decreased lactose catabolic process
 
|
 
|
 
Metabolic Activity
 
Metabolic Activity
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NCBI:562
 
NCBI:562
 
|
 
|
NG545-pi(1)<sub>b</sub>B
+
NG24
 
|
 
|
 
lacZ  
 
lacZ  
 
|
 
|
reduced acetylase activity
+
*Decreased acetylase activity
 +
*Decreased lactose catabolic process
 
|
 
|
 
Metabolic Activity
 
Metabolic Activity
 
|
 
|
33.5% of the wild type level when induced
+
4.7% of the wild type level when induced
 
|
 
|
 
Biochemical Assay
 
Biochemical Assay
 
|
 
|
Tables 3 and 4   
+
Table 4   
  
 
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{{RefHelp}}
 
{{RefHelp}}
 
<references/>
 
<references/>
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[[Category:Publication]]
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[[Category:To Be Converted]]

Latest revision as of 12:09, 22 June 2011

Citation

Grodzicker, T and Zipser, D (1968) A mutation which creates a new site for the re-initiation of polypeptide synthesis in the z gene of the lac operon of Escherichia coli.J. Mol. Biol. 38:305-14

Abstract

No abstract in PubMed

Links

PubMed

Keywords

Polar Effect; Acyltransferases; Chromosome Mapping; Escherichia coli; Genes; Membrane Transport Proteins; Mutation; Operon; Translation; Suppressor

Main Points of the Paper

  • Polar effects of a UAG mutation in the operator-proximal region of the z gene are studied which leads to the isolation of two suppressor mutations
  • π, a new site of initiation of protein synthesis is described
    • operates efficiently to restore permease and acetylase activity to a high level when induced
    • regulation/induction is still required, so the authors deduce that π is a new site of translation initiation (
    • not a new site of transcriptional initiation because acetylase is not present in the absence of inducer

Materials and Methods Used

  • Parent Strain: NG545 (trp-8, SmS Hfr)
    • UAG mutation had not been altered in the suppressor strains isolated
  • Strain: CA77B (Δlac, trp-8, SmR F-)
  • Strain: X103B (Δlac pro, trp-8, SmR F-)
  • Strain: M107 (Δlac, suI+, SmR F-)
  • Strain: ΔLacW (Δlac, his-, SmR F-)
  • Strain: E29 (lac+ , F'/lac+, threonine-, leucine-, B1-, SmR)
  • Acetylase assay
  • β-galactosidase assay

Phenotype Annotations

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

Species Taxon ID Strain Gene (if known) OMP Phenotype Details Evidence Notes

Escherichia coli

NCBI:562

NG545-pi(1)a

lacZ

  • Growth on melibiose medium
  • Melibiose catabolic process

Growth

growth on melibiose- polar effects

Plating Assay

Table 2

Escherichia coli

NCBI:562

NG545-pi(1)

lacZ

  • Suppressor-dependent growth on lactose medium
  • Suppressor-dependent lactose catabolic process

Growth

able to metabolize lactose only in a suI+ background

Plating Assay

Table 2- UAG polar mutation

Escherichia coli

NCBI:562

NG545-pi(1)b

lacZ

  • Abolished melibiose catabolic process
  • Absent growth on melibiose medium

Growth

unable to metabolize melibiose/ growth on melibiose-polar effects

Plating Assay

Table 2-

Escherichia coli

NCBI:562

NG545-pi(1)a

lacZ

  • Decreased gene expression of beta-galactosidase
  • Decreased lactose catabolic process

Metabolic Activity

trace amounts when induced

Biochemical Assay

Table 2

Escherichia coli

NCBI:562

NG545-pi(1)b

lacZ

reduced beta-galactosidase activity

Metabolic Activity

trace amounts when induced

Biochemical Assay

Table 2

Escherichia coli

NCBI:562

NG545B

lacZ

  • Decreased acetylase activity
  • Decreased lactose catabolic process

Metabolic Activity

4.2% of the wild type level when induced

Biochemical Assay

Tables 3 and 4

Escherichia coli

NCBI:562

NG545

lacZ

  • Suppressor-dependent growth on lactose medium
  • Suppressor-dependent lactose catabolic process

Growth

able to metabolize lactose only in a suI+ background

Plating Assay

Table 2- UAG polar mutation

Escherichia coli

NCBI:562

NG545-pi(1)

lacZ

  • Decreased acetylase activity
  • Decreased lactose catabolic process

Metabolic Activity

35% of the wild type level when induced

Biochemical Assay

Tables 3 and 4

Escherichia coli

NCBI:562

NG24

lacZ

  • Decreased acetylase activity
  • Decreased lactose catabolic process

Metabolic Activity

4.7% of the wild type level when induced

Biochemical Assay

Table 4

</protect>

Notes

References

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