Insertion of foreign random sequences of 120 amino acid residues into an active enzyme

Nobuhide Doi, Mitsuhiro Itaya, Tetsuya Yomo, Seiichi Tokura, Hiroshi Yanagawa

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Random sequences of 120-130 amino acid residues were inserted into a surface loop region of Escherichia coli RNase HI. This library was screened and about 10% of the clones were found to retain RNase H activity. Subsequent random mutagenesis led to an increase in RNase H activity and solubility of the protein. The inserted regions mere found not to contribute to the secondary structure of the mutant protein. The high frequency of insertion of flexible sequences and the increase in the protein's function by further mutagenesis simulate one of the events in protein evolution.

Original languageEnglish
Pages (from-to)177-180
Number of pages4
JournalFEBS Letters
Volume402
Issue number2-3
DOIs
Publication statusPublished - 1997 Jan 27

Keywords

  • Insertional mutagenesis
  • Protein evolution

ASJC Scopus subject areas

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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