Cloning and sequence analysis of cDNA encoding phosphatidic acid phosphatase (PAP) in coconut (Cocos nucifera L.).

Date

10-2005

Degree

Bachelor of Science in Biology

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Rita P. Laude

Abstract

Studies on the genes involved in the fatty acid synthesis (FAS) of coconut (Cocos inicifera L.) are essential in the improvement of the coconut oil production through genetic manipulation. One of the genes involved in coconut FAS is phosphatidic acid phosphatase (PAP). The cDNA of coconut PAP, synthesized using the 3'RACE technique, was cloned using pCR® 4-TOPO (lnvitrogen) vector. Plasmid DNA from the transformed Escherichia coli cells was purified using Perfectprep' Plasmid Mini (Eppendorf) and was sent to Macrogen sequencing facility in Seoul, South Korea. Sequence analysis indicates successful cloning of a partial coconut PAP gene isoforms. BLAST analysis showed significant similarities with known PAP including those from plants namely Oiyza sativa, Arabidopsis thaliana and Vigna unguiculata. A putative membrane bound protein from Bacteroides fragilis and the wunen protein from Drosophila melanogaster were also found to have significant similarity with the coconut PAP. Sequence motifs highly similar to conserved central and c-terminal phosphatases domains were identified in the PAP sequences suggesting that the cloned products are from the different regions of the coconut PAP gene. This study set the groundwork for full-length cloning of the coconut PAP gene.

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

Thesis

Document Type

Thesis

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