Isolation of Nucleotide Binding Site (NBS)-Leucine Rich Repeat (LRR) Resistant Gene Analogs (Rgas) In Arabica Coffee (Coffea Arabica L. Cv S.288)

• DOI: 10.4172/2155-952x.1000146
• Publication Date: 2012-09-28T09:09:47Z
• Authors: Deepak Kumar
• Keywords: genomic DNA; Amplicon

Cloning of resistance gene analogues against diverse pathogens from variety plants in last decade has revealed that many them share high level conserved sequence motifs. The backbone amino acid motifs present Nucleotide Binding Site (NBS) domain makes it possible to isolate by Polymerase chain reaction (PCR) with degenerate primers. Oligo-nucleotide primers combinations target conserve motif NBS as mentioned earlier studies were used amplify Coffea arabica (S.288). PCR product amplified genomic DNA well cDNA cloned and sequenced. In the study C.arabica using Ploop-cof GLPL-cof cloned, sequenced T7 / SP6 primers, analyzed at NCBI/SGN Data Bank. Analysis these RGA leads understanding difference expression profile might be due Resistant Gene Analogs (RGA) isolated cDNA. also presence similarity their sequences. Seven S.288 non-degenerate eleven more oligo-nucleotide thirty two S.288. Fifteen clones prepared rust race I infected leaf sample for 24 hours. BLASTN result showed a C.canephora RGA. This confirm integrity maintained among even was different coffee which there genome (C.arabica 2n= 44 22). below 475 bp or above 530 size along both primer sequences end either no match any they microsatellite. There is one independent four not given result, indicates may belong new type reported current are mainly class A