Poster: Plant-pathogen interactions
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Abs #
367: Isolation and characterization of NgRLK1, a receptor-like kinase of Nicotiana glutinosa that interacts with the elicitin of Phytophthora capsici
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Presenter: |
Kim, Yeong-Tae , Contact Author |
Authors | Kim, Yeong-Tae (A) Park, Jong-Sug (A) Kim, Kyung-Hwan (A) Kim, Jong-Bum (A) Go, Hyun-Jeong (A) Oh, Sang-Soo (A) Cho, Kang-Jin (A) | | Affiliations: |
(A): National Institute of Agricultural Biotechnology
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The capsicein was purified from culture filtrates of Phytophthora capsici by FPLC, and its partial primary structure was determined. The capsicein cDNA was cloned RT-PCR with degenerate primers based on conserved domains of known elicitins, which is composed of 294 nucleotides encoding 98 amino acids. The gene was cloned into pGEX 4T-1 vector and its expression was induced. As the gene product was applied to N. glutinosa, hypersensitive response in N. glutinosa was induced. Yeast two-hybrid technique was employed to identify potential protein interacting with capsicein. The two-hybrid library in yeast strain was screened with 5.7 x105 clones/library, and selected one (receptor-like kinase, RLK) homologous to serine/threonine kinase. The full-length clone of RLK was synthesized by 5'-rapid amplification of cDNA ends(RACE)-PCR, which was 2,749 bp in length and contained an ORF of 2,499 bp encoding 832 amino acids. The deduced polypeptide, designated NgRLK1, was molecular mass of 93,489Da and isoelectric point of 6.67 by the computational analysis. The structural feature of NgRLK1 shows the typical receptor-like kinases. It is composed of three main domains; a putative extracellular domain, a single transmembrane domain, and a C-terminal cytoplasmic serine/threonine protein kinase domain. NgRLK1 appears to be slightly induced by the capsicein treatment, based on RT-PCR analysis and the mRNA of NgRLK1 was existed intrinsically with low levels. Genomic Southern blot analysis revealed that NgRLK1 seems to be more than one copy gene in the N. glutinosa genome.
