Isolation and Sequence Analysis of LRR-RLK (Leucine-Rich Repeat Receptor-Like Protein Kinase) Gene of Kelampayan (Neolamarckia Cadamba)

Neolamarckia cadamba or locally known as kelampayan, belongs to the Rubiaceae family. N. cadamba is a fastgrowing tree species and has economic importance in timber industry. Leucine-rich repeat receptor-like protein kinase (LRR-RLK) genes are the largest group of receptor-like kinase (RLK) encoding genes. LRR-RLKs have been studied comprehensively for their essential roles in plant development and stress responses. This rises the needs to determine the gene sequence of LRR-RLK in N cadamba and the identity of the gene sequence as compared to the similar gene in other plant species. Total genomic DNA was extracted from the leaves of N. cadamba using cetyltrimethylammonium bromide (CTAB) followed by DNA purification and integrity assessment. Four EST sequences were retrieved from N. cadamba EST database for in silico analysis. Primers were designed based on comparison of LRR-RLK EST sequences of N. cadamba with genomic sequence of another Rubiaceae plant. Gradient polymerase chain reaction (PCR) was utilized to amplify the LRR-RLK partial gene and amplicons with desired size of approximately 619 bp were obtained. The PCR products of LRR-RLK partial gene was sequenced and in silico analysis on these sequences was conducted. This effort could provide data for tissuespecific expression model, for selection of tree genetic properties and functional analysis of LRR-RLK genes in N. cadamba