Isolation and characterisation of a NBS-LRR resistance gene analogue from pearl millet

Ranjini, P. ; Shailasree, S. ; Ramachandra Kini, K. ; Shekar Shetty, H. (2011) Isolation and characterisation of a NBS-LRR resistance gene analogue from pearl millet Archives of Phytopathology and Plant Protection, 44 (10). pp. 1014-1024. ISSN 0323-5408

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Official URL: http://www.tandfonline.com/doi/abs/10.1080/0323540...

Related URL: http://dx.doi.org/10.1080/03235401003633907

Abstract

Plant resistance (R) proteins belonging to nucleotide-binding site-leucine-rich repeat (NBS-LRR) family are mainly involved in recognition of effectors secreted by pathogens. Pearl millet [Pennisetum glaucum (L.) R.Br] is one of the most drought tolerant cereals, staple food crop of the semi-arid tropics but is highly susceptible to the downy mildew disease caused by oomycetous Sclerospora graminicola (Sacc) schroet. Earlier studies have identified several resistance gene analogues (RGAs) in pearl millet which may be involved in resistance against downy mildew. Of these, a clone RGPM213 was shown to have more than 60% identity with R-proteins coding for NBS-LRR-like protein kinase. The exact nature and function of the R-protein encoded by this gene was not known. In the present study, the cDNA of RGPM213 encompassing NBS-LRR region was inserted into an expression vector pRSET-A and transformed into BL21 E.coli cells. The expressed recombinant fusion protein with a His tag was purified using nickel affinity purification and it had a molecular weight of 35 kDa on SDS-PAGE. Immunoaffinity purification using antibodies raised against this recombinant R-protein identified two proteins of molecular weights 55 kDa and 66 kDa from pearl millet seedling extracts. Peptide mass fingerprinting of these proteins followed by homology search in database revealed similarity of the 55 kDa protein with a protein kinase from Brassica oleracia containing serine/ threonine kinase domain.

Item Type:Article
Source:Copyright of this article belongs to Taylor and Francis Group.
Keywords:Leucine Rich Repeat; Nucleotide Binding Site; Pennisetum glaucum; Resistance Gene Analogues(RGAs)
ID Code:48594
Deposited On:14 Jul 2011 14:11
Last Modified:14 Jul 2011 14:11

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