Isolation and sequence analysis of Cell-wall Associated Kinases (WAKs) Gene from Neolamarckia Cadamba

Siti Noor Azreen, Azli (2017) Isolation and sequence analysis of Cell-wall Associated Kinases (WAKs) Gene from Neolamarckia Cadamba. [Final Year Project Report] (Unpublished)

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Abstract

Neolamarckia cadamba is locally known as Kelampayan. It is used for industrial plantation and reforestation because of its fast growing properties. Cell wall-associated kinases (WAKs) are the functional proteins that are associated with the cell wall and required in the response to plant pathogen and also for cell expansion by activation from other environmental stimuli. Previous studies on WAKs are well developed in other species but there is a little research about WAK gene in N. cadamba. In this study, DNA was extracted from Ncadamba young leaves and subjected to Polymerase Chain Reaction (PCR) in order to amplify targeted gene using a previously designed primer set WAKI-F and WAKI-R. About -700 bp of amplicons were obtained and subjected to sequencing. BLASTn analysis was done to check for homology sequences and validate the data. Obtained gene were identified have high similarity with Nicotiana tabacum wall-associated kinase-like 20, mRNA about 72% similarity with E-value 3e-96. Phylogenetic tree that has been constructed showed that the isolated partial sequence of WAKs gene from N. cadamba was clustered in wall-associated kinases like 20 (WAKL20). By comparing with genomic DNA, mRNA and the obtain sequence it showed that there is no non-coding region was found in the sequence.

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