PCR-based detection of Acyl Carrier Protein (ACP) mRNA in various stages of coconut(Cocos nucifera L.) endosperm development.

Date

3-2003

Degree

Bachelor of Science in Biology

Major Course

Major in Cell Biology

College

College of Arts and Sciences (CAS)

Adviser/Committee Chair

Rita P. Laude

Co-adviser

Marni E. Cueno

Abstract

Detection of acyl carrier protein (ACP), a chloroplast-localized cofactor involved in the regulation of fatty acid biosynthesis, was done using mRNA isolated from 4-, S-and 6- month old (m.o.) coconut endosperm (?500 ng/pL). cDNAs were generated by using 3' RACE (Rapid Amplification of cDNA Ends) with two protocols, namely: (1) the protocol for conventional transcripts; and (2) the protocol for GC rich transcripts. No products were produced using the protocol for conventional transcripts. cDNAs generated using the protocol for GC rich transcripts have concentrations of 0.1 ng/uL, 0.1-0.5 ng/pL, 0.5-1.0 ng/uL in 4-, 5- and 6-m.o. coconut endosperms, respectively. It is assumed that coconut ACP gene has high GC content since no products were produced using the conventional protocol. Using cDNAs synthesized from the second protocol, COLD-START PCR produced a single electrophoretic band of 300 by in only 4-m.o. coconut endosperm. No bands were produced in 5- and 6-m.o. coconut endosperms. This may suggest that the acyl carrier protein is only expressed in 4-m.o. coconut endosperm but not in 5- and 6-m.o. coconut endosperms. No isoforms were detected in 4-m.o. coconut endosperm probably due to the tissue-specific expression of ACP.

Language

English

Location

UPLB Main Library Special Collections Section (USCS)

Call Number

Thesis

Document Type

Thesis

This document is currently not available here.

Share

COinS