Designing and Testing of Primers for Isolation of Expansin Gene from Canarium Odontophyllum (Miq.) Dabai

Harttini Neeni, Hatta (2008) Designing and Testing of Primers for Isolation of Expansin Gene from Canarium Odontophyllum (Miq.) Dabai. [Final Year Project Report] (Unpublished)

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Abstract

Canarium odontophyllum which is locally known as dabai, is one of important indigenous wild fruit species in Sarawak. These exotic fruits are found in several divisions such as Sibu, Kapit and Sarikei. These varieties exhibit different thickness of mesocarp. This characteristic is regulated by many complex genes and one of the key genes involved is the expansin genes. These genes function in cell wall expansion that contributes to the thickness of mesocarp. Thus, this study focuses on designing and testing primers for isolation of expansin genes from C.odontophyllum. Advance understanding in expansin genes can be applied in crop improvement to cultivate C.odontophyllum in commercial scale and to produce varieties that have better traits and higher market demand. Total genomic DNA was isolated successfully using modified CT AB methods described by Doyle and Doyle (1990). A pair of primers for EXP A gene that regulate cell enlargement was designed and synthesized based on the nucleotide sequence from Populus tremula x Populus tremuloides alpha-expansin 1. These primers were used for amplification of EXPA from the genomic DNA of C.odontophyllum. Various annealing temperatures were used to optimize the peR product obtained. Other parameters such as the concentration of MgCI 2, concentration of DNA template and concentration of primers were optimized for attaining optimum PCR products.

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