Heterologous expression of recombinant alcohol dehydrogenase gene from sago palm in bacterial and plant system

Alcohol dehydrogenase (ADH) is an enzyme involved in pathways that respond to various environmental stresses such as osmotic, wound and anaerobic condition. In this study, the recombinant Metroxylon sagu Adh1 cDNA (r-msAdh1) was cloned into an expression vector; pET-41a(+) and expressed in Escherich...

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Main Author:	Mastura, Sani
Format:	Thesis
Language:	English
Published:	2016
Subjects:	TP Chemical technology
Online Access:	http://ir.unimas.my/id/eprint/30641/1/Mastura%20Sani%20ft.pdf
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id	my-unimas-ir.30641
record_format	uketd_dc
spelling	my-unimas-ir.306412023-10-03T04:15:49Z Heterologous expression of recombinant alcohol dehydrogenase gene from sago palm in bacterial and plant system 2016 Mastura, Sani TP Chemical technology Alcohol dehydrogenase (ADH) is an enzyme involved in pathways that respond to various environmental stresses such as osmotic, wound and anaerobic condition. In this study, the recombinant Metroxylon sagu Adh1 cDNA (r-msAdh1) was cloned into an expression vector; pET-41a(+) and expressed in Escherichia coli (E. coli) strain BL21 (DE3). SDS-PAGE analysis of the E. coli lysate revealed that large amount of expressed protein formed insoluble aggregation, non-active protein. However, soluble r-msAdh1 protein was successfully produced at low-growth temperature and showed catalytic activity when spectrophotometrically assayed. Second part of this study involves the in-planta transformation of r-msAdh1 cDNA into tomato seeds using Agrobacterium tumefaciens (A. tumefaciens) strain LBA4404. The result showed that, out of 15 putative transformed plants (T0) were analysed, five putative transgenic plant lines were determined to carry r-msAdh1 cDNA in their genome. Further analysis also showed that r-msAdh1 cDNA have passed into all T1 transgenic lines and expression analysis at transcript level confirmed the presence of r-msAdh1 in transgenic tomato genome. Universiti Malaysia Sarawak(UNIMAS) 2016 Thesis http://ir.unimas.my/id/eprint/30641/ http://ir.unimas.my/id/eprint/30641/1/Mastura%20Sani%20ft.pdf text en validuser masters Universiti Malaysia Sarawak(UNIMAS) Faculty of Resource Science and Technology
institution	Universiti Malaysia Sarawak
collection	UNIMAS Institutional Repository
language	English
topic	TP Chemical technology
spellingShingle	TP Chemical technology Mastura, Sani Heterologous expression of recombinant alcohol dehydrogenase gene from sago palm in bacterial and plant system
description	Alcohol dehydrogenase (ADH) is an enzyme involved in pathways that respond to various environmental stresses such as osmotic, wound and anaerobic condition. In this study, the recombinant Metroxylon sagu Adh1 cDNA (r-msAdh1) was cloned into an expression vector; pET-41a(+) and expressed in Escherichia coli (E. coli) strain BL21 (DE3). SDS-PAGE analysis of the E. coli lysate revealed that large amount of expressed protein formed insoluble aggregation, non-active protein. However, soluble r-msAdh1 protein was successfully produced at low-growth temperature and showed catalytic activity when spectrophotometrically assayed. Second part of this study involves the in-planta transformation of r-msAdh1 cDNA into tomato seeds using Agrobacterium tumefaciens (A. tumefaciens) strain LBA4404. The result showed that, out of 15 putative transformed plants (T0) were analysed, five putative transgenic plant lines were determined to carry r-msAdh1 cDNA in their genome. Further analysis also showed that r-msAdh1 cDNA have passed into all T1 transgenic lines and expression analysis at transcript level confirmed the presence of r-msAdh1 in transgenic tomato genome.
format	Thesis
qualification_level	Master's degree
author	Mastura, Sani
author_facet	Mastura, Sani
author_sort	Mastura, Sani
title	Heterologous expression of recombinant alcohol dehydrogenase gene from sago palm in bacterial and plant system
title_short	Heterologous expression of recombinant alcohol dehydrogenase gene from sago palm in bacterial and plant system
title_full	Heterologous expression of recombinant alcohol dehydrogenase gene from sago palm in bacterial and plant system
title_fullStr	Heterologous expression of recombinant alcohol dehydrogenase gene from sago palm in bacterial and plant system
title_full_unstemmed	Heterologous expression of recombinant alcohol dehydrogenase gene from sago palm in bacterial and plant system
title_sort	heterologous expression of recombinant alcohol dehydrogenase gene from sago palm in bacterial and plant system
granting_institution	Universiti Malaysia Sarawak(UNIMAS)
granting_department	Faculty of Resource Science and Technology
publishDate	2016
url	http://ir.unimas.my/id/eprint/30641/1/Mastura%20Sani%20ft.pdf
_version_	1783728389784338432

Heterologous expression of recombinant alcohol dehydrogenase gene from sago palm in bacterial and plant system

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