Heterologous Expression of the Avirulence Gene Product, NIP1, from the Barley Pathogen Rhynchosporium secalis

NIP1, the product of the avirulence gene AvrRrs1 from Rhynchosporium secalis, a fungal pathogen of barley, is a small secreted cysteine-rich protein. This protein is essential for the specific recognition of the fungus by host plants carrying the complementary resistance gene Rrs1. Different heterol...

Full description

Saved in:
Bibliographic Details
Published in:Protein expression and purification 1999-10, Vol.17 (1), p.64-73
Main Authors: Gierlich, Angela, van 't Slot, Klaas A.E., Li, Volkhart M., Marie, Corinne, Hermann, Hanno, Knogge, Wolfgang
Format: Article
Language:English
Subjects:
Animals
Ascomycota - genetics
Ascomycota - pathogenicity
Baculoviridae - genetics
Base Sequence
Cell Line
DNA Primers - genetics
Escherichia coli - genetics
Eukaryotic Initiation Factor-3
Fungal Proteins - biosynthesis
Fungal Proteins - genetics
Fungal Proteins - isolation & purification
Gene Expression
Genes, Fungal
Hordeum - microbiology
Molecular Weight
Nuclear Proteins - biosynthesis
Nuclear Proteins - genetics
Nuclear Proteins - isolation & purification
Pichia - genetics
Recombinant Proteins - biosynthesis
Recombinant Proteins - genetics
Recombinant Proteins - isolation & purification
Saccharomyces cerevisiae Proteins
Spodoptera
Virulence - genetics
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:NIP1, the product of the avirulence gene AvrRrs1 from Rhynchosporium secalis, a fungal pathogen of barley, is a small secreted cysteine-rich protein. This protein is essential for the specific recognition of the fungus by host plants carrying the complementary resistance gene Rrs1. Different heterologous expression systems were tested to produce sufficient quantities of NIP1 to allow its utilization in receptor identification and isolation. In addition, protein amounts higher than those produced in fungal cultures are required to determine its 3D structure and to analyze its interaction with a receptor. The most efficient method, the synthesis of a His-tag fusion protein in Escherichia coli combined with a refolding procedure, yielded up to 3 mg of recombinant NIP1 from a 1-liter bacterial culture. After removal of the His-tag, the recombinant protein showed the same physicochemical characteristics as the native NIP1 and, most importantly, full biological activity.
ISSN:1046-5928
1096-0279
DOI:10.1006/prep.1999.1098