Expression of nicotiana glutinosa ribonucleases in escherichia coli

Madoka Hino, Shin Kawano, Makoto Kimura

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Abstract

We previously isolated from Nicotiana glutinosa leaves three distinct cDNA clones, NGR1, NGR2, and NGR3, encoding a wound-inducible RNase NW, and putative RNases NGR2 and NGR3, respectively. In this study, we produced RNases NW and NGR3 in Escherichia coli and purified them to homogeneity. RNase NGR3 had non-absolute specificity toward polynucleotides, although RNase NW preferentially cleaved polyinosinic acid (Poly I). Both RNases NW and NGR3 were more active toward diribonucleoside monophosphates ApG, CpU, and GpU. Furthermore, kinetic parameters for RNase NW (Km, 0.778 mM and kcat, 1938 min−1) and RNase NGR3 (Km, 0.548 mM and kcat, 408 min−1) were calculated using GpU as a substrate.

Original languageEnglish
Pages (from-to)910-912
Number of pages3
JournalBioscience, Biotechnology and Biochemistry
Volume66
Issue number4
DOIs
Publication statusPublished - Jan 1 2002

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All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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