Plant, Animal & Microbe Genomes X Conference Plant, Animal & Microbe Genomes X ConferenceJanuary 12-16, 2002
Town & Country Convention Center
San Diego, CA
Poster: Genome Sequencing & ESTs
Motifs are assigned for specific functions among a protein family, and are widely used to identify functional regions of proteins, their ancestors and family classifications. P-loop (kinase-1a) is frequently occurred and shared motif by several nucleotide-binding site encoding (NBS-encoding) proteins. We used NBS-specific PCR primers and utilized a modified polyacrylamide gel by the cDNA technology to clone new NBS-encoding genes, by screening whole-wheat genome. With several 3' to 5' T-oligonucleotide primers, we used an NBS-I type primer encoding p-loop (kinase-1a; GVGKTT) and a hydrophobic motif encoding GLPLAL. We inoculated resistant wheat lines with their corresponding pathogens to clone induced gene members along with constitutive members. So far, we cloned 350 fragments, which were categorized in 121 classes. Majority of the fragments were found to belong to NBS-encoding proteins including protein kinases, serin/threonine kinases, receptor like protein kinases, NBS-LRRs class of resistance genes and some other stress related genes. According to the blast search results, 37 classes were assigned to either a definite (80% and over protein sequence identity) or a putative function (35% to 80% identities). Motif search among unknown classes resulted in 18 new NBS-LRR type of resistance and/or apoptosis related-like genes containing NBS motifs (kinase-1a, kinase-2 and kinase-3a). Sixty-one fragments remain unknown even though 45 of these contain possible NBS motifs. Virtual northern in public non-redundant (nr) and expressed tag sequence (EST) database (www.ncbi.nih.nlmn.gov) resulted in highly expressed 19 fragments (16.4%) of cloned genes (present in the database more than 2 transcript), however, 102 fragments (83.8 %) have been very rare and/or not present in any present EST libraries.