Abstract:
: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.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Gierlich A,van 't Slot KA,Li VM,Marie C,Hermann H,Knogge Wdoi
10.1006/prep.1999.1098keywords:
subject
Has Abstractpub_date
1999-10-01 00:00:00pages
64-73issue
1eissn
1046-5928issn
1096-0279pii
S1046-5928(99)91098-7journal_volume
17pub_type
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