Isoforms of phosphatidic acid phosphatase (PAP) gene in coconut (Cocos nucifera L.)

Date

3-2003

Degree

Bachelor of Science in Biology

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Rita P. Laude

Abstract

The isoforms of phosphatidic acid phosphatase (PAP) gene involved in the fatty acid synthesis in coconuts were determined using the 3' rapid amplification of cDN ends (3' RACE) technique. The mRNAs of the 4-, 5-, and 6-month old coconendosperm tissues were extracted and synthesized to cDNAs by reverse transcriptase. Amplification of cDNAs were performed using three (3) types of polymerase chain reaction (PCR): cold-start and combined hot-start and touchdown. Electrophoretic analysis of the 3'RACE products using cold-start PCR did not show bands in the 4-, 5-,and 6-month-old samples. This may be due to the formation of secondary structures caused by the long size of the PAP gene or the GC rich sequence of the coconut which was already previously noted in another study. This, therefore. required a longer denaturing time for the cDNAs. Electrophoretic analysis of the 3'RACE products using the combined hot-start and touchdown PCR showed three (3) isoforms in the 6-month-old only which were 400 bp, 500 bp. and 700 by in length. However, no isoforms were observed in the 4- and 5-month-old coconuts. This suggests that while the enzyme is not yet needed by the endosperm for oil accumulation at the 4- and 5-month-old. at 6-month-old rapid oil accumulation occurs; hence its expression. Furthermore, PAP activity is "developmental)} induced". Expression of the gene increases as the coconut matures.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

Thesis

Document Type

Thesis

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