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T41 mutation in lac repressor is Tyr282----Asp.

J Chen1, K S Matthews

  • 1Department of Biochemistry and Cell Biology, Rice University, Houston, TX 77251.

Gene
|February 1, 1992
PubMed
Summary

A novel lac repressor mutation, T41, was identified as a Tyr282Asp substitution. This mutation alters the protein

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Area of Science:

  • Molecular biology
  • Protein engineering
  • Genetics

Background:

  • The lac repressor protein controls gene expression in Escherichia coli.
  • Previous studies identified mutations at Tyr282 affecting repressor function.
  • The T41 mutant protein was previously characterized as monomeric.

Purpose of the Study:

  • To identify the specific mutation responsible for the T41 phenotype.
  • To characterize the biochemical properties of the T41 mutant protein.

Main Methods:

  • Polymerase chain reaction (PCR) amplification and sequencing of the T41 gene.
  • Site-specific mutagenesis to create the Tyr282Asp mutant.
  • Isolation and characterization of the mutant protein.

Main Results:

  • The T41 mutation was precisely mapped to a Tyr282Asp amino acid substitution.
  • Site-specific production of the Tyr282Asp mutant confirmed its properties.
  • The Tyr282Asp mutant protein exhibits monomeric and altered inducer binding characteristics.

Conclusions:

  • The Tyr282Asp substitution is responsible for the observed T41 mutant phenotype.
  • This finding provides a precise molecular basis for altered lac repressor function.
  • The study validates the role of residue 282 in repressor oligomerization and ligand binding.

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