Characterisation and Functional Analysis of Isochorismate Synthase cDNA from Morinda citrifolia L.

Isochorismate synthase catalyzes the substrate chorismate in the presence of a metal ion (Mg2+) cofactor to isochorismate. Isochorismate is prominent in the shikimate pathway to produce siderophores, anthraquinones and salicylate. In previous study, a near complete cDNA encoding isochorismate synth...

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Main Author:	Chai, Suk Phin
Format:	Thesis
Language:	English
Published:	2016
Subjects:	TP Chemical technology
Online Access:	http://ir.unimas.my/id/eprint/30586/1/Chai%20Suk%20Phin%20ft.pdf
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id	my-unimas-ir.30586
record_format	uketd_dc
spelling	my-unimas-ir.305862023-04-27T03:18:59Z Characterisation and Functional Analysis of Isochorismate Synthase cDNA from Morinda citrifolia L. 2016 Chai, Suk Phin TP Chemical technology Isochorismate synthase catalyzes the substrate chorismate in the presence of a metal ion (Mg2+) cofactor to isochorismate. Isochorismate is prominent in the shikimate pathway to produce siderophores, anthraquinones and salicylate. In previous study, a near complete cDNA encoding isochorismate synthase (mcICS) has been isolated from ‘mengkudu’ or Morinda citrifolia. The cDNA was characterized and identified to be highest identity of 72% to Catharanthus roseus (AJ006065). An in silico analysis of mcICS cDNA was studied in this research to determine the 3D structure and active amino acid residues. The mcICS cDNA was further analyzed by cloning into pET-41a(+) vector for expression analysis in prokaryote system. pET-41a(+)/mcICS was constructed and yielded molecular weight approximately 55kDa. The mcICS cDNA also cloned into pGSA1131 vector and transformed into mungbean (Vigna radiata) through a direct cotyledons Agrobacterium tumefaciens- mediated transformation. This type of transformation was less laborious,efficient and it showed 3.33 % of transformation efficiency. The putative transgenic were grown to maturity to analyse the stability of transgene integration and expression of T1 generation. Results showed that some individulas from T1 generation as indicated by posttransciptional gene siliencing which observed as chimera as the transgenic mungbean construct from different cells of zygotes. Hence, these results suggests to analyze next T2 generation to provides more stability for transgenes integration and expression. Universiti Malaysia Sarawak(UNIMAS) 2016 Thesis http://ir.unimas.my/id/eprint/30586/ http://ir.unimas.my/id/eprint/30586/1/Chai%20Suk%20Phin%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 Chai, Suk Phin Characterisation and Functional Analysis of Isochorismate Synthase cDNA from Morinda citrifolia L.
description	Isochorismate synthase catalyzes the substrate chorismate in the presence of a metal ion (Mg2+) cofactor to isochorismate. Isochorismate is prominent in the shikimate pathway to produce siderophores, anthraquinones and salicylate. In previous study, a near complete cDNA encoding isochorismate synthase (mcICS) has been isolated from ‘mengkudu’ or Morinda citrifolia. The cDNA was characterized and identified to be highest identity of 72% to Catharanthus roseus (AJ006065). An in silico analysis of mcICS cDNA was studied in this research to determine the 3D structure and active amino acid residues. The mcICS cDNA was further analyzed by cloning into pET-41a(+) vector for expression analysis in prokaryote system. pET-41a(+)/mcICS was constructed and yielded molecular weight approximately 55kDa. The mcICS cDNA also cloned into pGSA1131 vector and transformed into mungbean (Vigna radiata) through a direct cotyledons Agrobacterium tumefaciens- mediated transformation. This type of transformation was less laborious,efficient and it showed 3.33 % of transformation efficiency. The putative transgenic were grown to maturity to analyse the stability of transgene integration and expression of T1 generation. Results showed that some individulas from T1 generation as indicated by posttransciptional gene siliencing which observed as chimera as the transgenic mungbean construct from different cells of zygotes. Hence, these results suggests to analyze next T2 generation to provides more stability for transgenes integration and expression.
format	Thesis
qualification_level	Master's degree
author	Chai, Suk Phin
author_facet	Chai, Suk Phin
author_sort	Chai, Suk Phin
title	Characterisation and Functional Analysis of Isochorismate Synthase cDNA from Morinda citrifolia L.
title_short	Characterisation and Functional Analysis of Isochorismate Synthase cDNA from Morinda citrifolia L.
title_full	Characterisation and Functional Analysis of Isochorismate Synthase cDNA from Morinda citrifolia L.
title_fullStr	Characterisation and Functional Analysis of Isochorismate Synthase cDNA from Morinda citrifolia L.
title_full_unstemmed	Characterisation and Functional Analysis of Isochorismate Synthase cDNA from Morinda citrifolia L.
title_sort	characterisation and functional analysis of isochorismate synthase cdna from morinda citrifolia l.
granting_institution	Universiti Malaysia Sarawak(UNIMAS)
granting_department	Faculty of Resource Science and Technology
publishDate	2016
url	http://ir.unimas.my/id/eprint/30586/1/Chai%20Suk%20Phin%20ft.pdf
_version_	1783728388644536320

Characterisation and Functional Analysis of Isochorismate Synthase cDNA from Morinda citrifolia L.

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