Generation Of Expressed Sequence Tags (Ests) From Non-Embryogenic Callus Cdna Libraries Of Oil Palm (Elaeis Guineensis Jacq.)

Due to the economic importance of tissue culture in oil palm, it is desirable to understand the chemical characteristics and molecular changes associated with callogenesis and embryogenesis. Therefore partial sequencing of randomly selected complementary DNA (cDNA) clones was carried out to inves...

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Main Author:	Tan, Amos Chi Yee
Format:	Thesis
Language:	English English
Published:	2003
Subjects:	Oil palm - Embryology. Molecular cloning.
Online Access:	http://psasir.upm.edu.my/id/eprint/11862/1/FSMB_2003_10.pdf
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id	my-upm-ir.11862
record_format	uketd_dc
spelling	my-upm-ir.118622024-06-27T01:39:27Z Generation Of Expressed Sequence Tags (Ests) From Non-Embryogenic Callus Cdna Libraries Of Oil Palm (Elaeis Guineensis Jacq.) 2003-07 Tan, Amos Chi Yee Due to the economic importance of tissue culture in oil palm, it is desirable to understand the chemical characteristics and molecular changes associated with callogenesis and embryogenesis. Therefore partial sequencing of randomly selected complementary DNA (cDNA) clones was carried out to investigate and understand the gene expression pattern during non-embryogenic callus in oil palm. A total of 2,296 clones from cDNA libraries constructed using messenger RNA (mRNA) from nonembryogenic callus tissues have been partially sequenced to generate expressed sequence tags (ESTs). The comparison of the coding capacity of these 1 ,832 ESTs with the nonredundant sequences in the National Center for Biotechnology Information (NCBI) databases resulted in assignment of putative function to 971 (53.0%) of the ESTs. The remaining 861 (47.0%) were considered as novel genes. Hybridization of the 80 selected cDNA clones to 32P-Iabelled first strand cDNA probes synthesized from the total mRNA of NEC was compared with probes prepared from mRNA of leaves. A total of 1 2 clones, which hybridized predominantly to the NEC but not to the leaf probe were selected for Northern analysis.The Northern analysis showed that 9 out of the 12 ESTs clones selected were expressed predominantly in the callus tissue. However, carrying out a sequence comparison using the BLAST algorithm, only l out of the 9 clones had significant homology to a known gene, glutathione-S-transferase (GST) gene. The remaining 8 unidentifiable ESTs were then subjected to a motif search using the Blocks IMProved Searcher (BLIMPS) program. Based on the search results, 4 different motifs were identified to be related to the 8 ESTs and they are either involved in developmental regulation or protein storage function. The results presented here therefore demonstrate the effectiveness and efficiency of the EST strategy in gene discovery in the oil palm callus tissue. Oil palm - Embryology. Molecular cloning. 2003-07 Thesis http://psasir.upm.edu.my/id/eprint/11862/ http://psasir.upm.edu.my/id/eprint/11862/1/FSMB_2003_10.pdf text en public masters Universiti Putra Malaysia Oil palm - Embryology. Molecular cloning. Faculty of Food Science and Biotechnology Tan, Siang Hee English
institution	Universiti Putra Malaysia
collection	PSAS Institutional Repository
language	English English
advisor	Tan, Siang Hee
topic	Oil palm - Embryology. Molecular cloning.
spellingShingle	Oil palm - Embryology. Molecular cloning. Tan, Amos Chi Yee Generation Of Expressed Sequence Tags (Ests) From Non-Embryogenic Callus Cdna Libraries Of Oil Palm (Elaeis Guineensis Jacq.)
description	Due to the economic importance of tissue culture in oil palm, it is desirable to understand the chemical characteristics and molecular changes associated with callogenesis and embryogenesis. Therefore partial sequencing of randomly selected complementary DNA (cDNA) clones was carried out to investigate and understand the gene expression pattern during non-embryogenic callus in oil palm. A total of 2,296 clones from cDNA libraries constructed using messenger RNA (mRNA) from nonembryogenic callus tissues have been partially sequenced to generate expressed sequence tags (ESTs). The comparison of the coding capacity of these 1 ,832 ESTs with the nonredundant sequences in the National Center for Biotechnology Information (NCBI) databases resulted in assignment of putative function to 971 (53.0%) of the ESTs. The remaining 861 (47.0%) were considered as novel genes. Hybridization of the 80 selected cDNA clones to 32P-Iabelled first strand cDNA probes synthesized from the total mRNA of NEC was compared with probes prepared from mRNA of leaves. A total of 1 2 clones, which hybridized predominantly to the NEC but not to the leaf probe were selected for Northern analysis.The Northern analysis showed that 9 out of the 12 ESTs clones selected were expressed predominantly in the callus tissue. However, carrying out a sequence comparison using the BLAST algorithm, only l out of the 9 clones had significant homology to a known gene, glutathione-S-transferase (GST) gene. The remaining 8 unidentifiable ESTs were then subjected to a motif search using the Blocks IMProved Searcher (BLIMPS) program. Based on the search results, 4 different motifs were identified to be related to the 8 ESTs and they are either involved in developmental regulation or protein storage function. The results presented here therefore demonstrate the effectiveness and efficiency of the EST strategy in gene discovery in the oil palm callus tissue.
format	Thesis
qualification_level	Master's degree
author	Tan, Amos Chi Yee
author_facet	Tan, Amos Chi Yee
author_sort	Tan, Amos Chi Yee
title	Generation Of Expressed Sequence Tags (Ests) From Non-Embryogenic Callus Cdna Libraries Of Oil Palm (Elaeis Guineensis Jacq.)
title_short	Generation Of Expressed Sequence Tags (Ests) From Non-Embryogenic Callus Cdna Libraries Of Oil Palm (Elaeis Guineensis Jacq.)
title_full	Generation Of Expressed Sequence Tags (Ests) From Non-Embryogenic Callus Cdna Libraries Of Oil Palm (Elaeis Guineensis Jacq.)
title_fullStr	Generation Of Expressed Sequence Tags (Ests) From Non-Embryogenic Callus Cdna Libraries Of Oil Palm (Elaeis Guineensis Jacq.)
title_full_unstemmed	Generation Of Expressed Sequence Tags (Ests) From Non-Embryogenic Callus Cdna Libraries Of Oil Palm (Elaeis Guineensis Jacq.)
title_sort	generation of expressed sequence tags (ests) from non-embryogenic callus cdna libraries of oil palm (elaeis guineensis jacq.)
granting_institution	Universiti Putra Malaysia
granting_department	Faculty of Food Science and Biotechnology
publishDate	2003
url	http://psasir.upm.edu.my/id/eprint/11862/1/FSMB_2003_10.pdf
_version_	1804888661837217792

Generation Of Expressed Sequence Tags (Ests) From Non-Embryogenic Callus Cdna Libraries Of Oil Palm (Elaeis Guineensis Jacq.)

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