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A Transient Assay to Evaluate the Expression of Polyhydroxybutyrate Genes Regulated by Oil Palm Mesocarp-Specific Promoter

The promoter of the oil palm metallothionein-like gene (MT3-A) demonstrated mesocarp-specific activity in functional analysis using transient expression assay of reporter gene in bombarded oil palm tissue slices. In order to investigate the tissue-specific expression of polyhydroxybutyrate (PHB) bio...

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Main Authors: Omidvar, Vahid, Abdullah, Siti Nor Akmar, Mahmood, Maziah, Abdul Aziz, Maheran
Format: Article
Language:English
Published: Springer Verlag 2008
Online Access:http://psasir.upm.edu.my/id/eprint/857/
http://dx.doi.org/10.1007/s00299-008-0565-2
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spelling my.upm.eprints.8572011-10-30T05:11:23Z http://psasir.upm.edu.my/id/eprint/857/ A Transient Assay to Evaluate the Expression of Polyhydroxybutyrate Genes Regulated by Oil Palm Mesocarp-Specific Promoter Omidvar, Vahid Abdullah, Siti Nor Akmar Mahmood, Maziah Abdul Aziz, Maheran The promoter of the oil palm metallothionein-like gene (MT3-A) demonstrated mesocarp-specific activity in functional analysis using transient expression assay of reporter gene in bombarded oil palm tissue slices. In order to investigate the tissue-specific expression of polyhydroxybutyrate (PHB) biosynthetic pathway genes, a multi-gene construct carrying PHB genes fused to the oil palm MT3-A promoter was co-transferred with a construct carrying GFP reporter gene using microprojectile bombardment targeting the mesocarp and leaf tissues of the oil palm. Transcriptional analysis using RT-PCR revealed successful transcription of all the three phbA, phbB, and phbC genes in transiently transformed mesocarp but not in transiently transformed leaf tissues. Furthermore, all the three expected sizes of PHB-encoded protein products were only detected in transiently transformed mesocarp tissues on a silver stained polyacrylamide gel. Western blot analysis using polyclonal antibody specific for phbB product confirmed successful translation of phbB mRNA transcript into protein product. This study provided valuable information, supporting the future engineering of PHB-producing transgenic palms. Springer Verlag 2008 Article PeerReviewed Omidvar, Vahid and Abdullah, Siti Nor Akmar and Mahmood, Maziah and Abdul Aziz, Maheran (2008) A Transient Assay to Evaluate the Expression of Polyhydroxybutyrate Genes Regulated by Oil Palm Mesocarp-Specific Promoter. Plant Cell Reports, 27 (9). pp. 1451-1459. ISSN 0721-7714 http://dx.doi.org/10.1007/s00299-008-0565-2 English
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
description The promoter of the oil palm metallothionein-like gene (MT3-A) demonstrated mesocarp-specific activity in functional analysis using transient expression assay of reporter gene in bombarded oil palm tissue slices. In order to investigate the tissue-specific expression of polyhydroxybutyrate (PHB) biosynthetic pathway genes, a multi-gene construct carrying PHB genes fused to the oil palm MT3-A promoter was co-transferred with a construct carrying GFP reporter gene using microprojectile bombardment targeting the mesocarp and leaf tissues of the oil palm. Transcriptional analysis using RT-PCR revealed successful transcription of all the three phbA, phbB, and phbC genes in transiently transformed mesocarp but not in transiently transformed leaf tissues. Furthermore, all the three expected sizes of PHB-encoded protein products were only detected in transiently transformed mesocarp tissues on a silver stained polyacrylamide gel. Western blot analysis using polyclonal antibody specific for phbB product confirmed successful translation of phbB mRNA transcript into protein product. This study provided valuable information, supporting the future engineering of PHB-producing transgenic palms.
format Article
author Omidvar, Vahid
Abdullah, Siti Nor Akmar
Mahmood, Maziah
Abdul Aziz, Maheran
spellingShingle Omidvar, Vahid
Abdullah, Siti Nor Akmar
Mahmood, Maziah
Abdul Aziz, Maheran
A Transient Assay to Evaluate the Expression of Polyhydroxybutyrate Genes Regulated by Oil Palm Mesocarp-Specific Promoter
author_facet Omidvar, Vahid
Abdullah, Siti Nor Akmar
Mahmood, Maziah
Abdul Aziz, Maheran
author_sort Omidvar, Vahid
title A Transient Assay to Evaluate the Expression of Polyhydroxybutyrate Genes Regulated by Oil Palm Mesocarp-Specific Promoter
title_short A Transient Assay to Evaluate the Expression of Polyhydroxybutyrate Genes Regulated by Oil Palm Mesocarp-Specific Promoter
title_full A Transient Assay to Evaluate the Expression of Polyhydroxybutyrate Genes Regulated by Oil Palm Mesocarp-Specific Promoter
title_fullStr A Transient Assay to Evaluate the Expression of Polyhydroxybutyrate Genes Regulated by Oil Palm Mesocarp-Specific Promoter
title_full_unstemmed A Transient Assay to Evaluate the Expression of Polyhydroxybutyrate Genes Regulated by Oil Palm Mesocarp-Specific Promoter
title_sort transient assay to evaluate the expression of polyhydroxybutyrate genes regulated by oil palm mesocarp-specific promoter
publisher Springer Verlag
publishDate 2008
url http://psasir.upm.edu.my/id/eprint/857/
http://dx.doi.org/10.1007/s00299-008-0565-2
_version_ 1643821925761286144
score 13.211869

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