A genetic system for isolation and characterization of TaqI restriction endonuclease mutants. Academic Article uri icon

Overview

abstract

  • The gene encoding TaqI restriction endonuclease has been subcloned downstream from an inducible phoA promoter. Certain strains of Escherichia coli remain viable when endonuclease is expressed, even in the absence of (protective) methylation. Infecting lambda phage DNA is not restricted in vivo. One E. coli strain, MM294, exhibited a temperature-sensitive phenotype when TaqI endonuclease was induced. This allowed for design of an in vivo plate assay for identification of specially constructed two-codon insertion mutants in the endonuclease gene. These mutants exhibited a wide range of in vitro activities, including wild-type activity, greater activity in low-salt buffer, and sequence-specific nicking activity.

publication date

  • January 1, 1987

Research

keywords

  • DNA Restriction Enzymes
  • Deoxyribonucleases, Type II Site-Specific

Identity

Scopus Document Identifier

  • 0023464709

Digital Object Identifier (DOI)

  • 10.1016/0378-1119(87)90154-5

PubMed ID

  • 2824288

Additional Document Info

volume

  • 56

issue

  • 1