$Functional analysis of a latex-specific promoter (hevein) of Hevea for enhanced recombinant protein production in Hevea brasiliensis$

Functional analysis of a latex-specific promoter (hevein) of Hevea for enhanced recombinant protein production in Hevea brasiliensis

Any protein, either naturally occurring or designed synthetically, could be produced safely, inexpensively and in large quantities using the power of the rubber trees for the production of recombinant proteins on a plantation scale with sunlight and water. In the past, the Rubber Research Institu...

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Main Authors:	Pappusamy, Arokiaraj / PA, Shuib, Siti Shuhada, Enamalai, Sunderasan
Format:	Conference or Workshop Item
Language:	English
Published:	2011
Subjects:	SB Plant culture
Online Access:	http://irep.iium.edu.my/2759/ http://irep.iium.edu.my/2759/1/Arokiaraj_Pappusamy.22nd_Science_Congress_Conference_paper.2011.docx

id	iium-2759
recordtype	eprints
spelling	iium-27592011-12-15T13:30:59Z http://irep.iium.edu.my/2759/ Functional analysis of a latex-specific promoter (hevein) of Hevea for enhanced recombinant protein production in Hevea brasiliensis Pappusamy, Arokiaraj / PA Shuib, Siti Shuhada Enamalai, Sunderasan SB Plant culture Any protein, either naturally occurring or designed synthetically, could be produced safely, inexpensively and in large quantities using the power of the rubber trees for the production of recombinant proteins on a plantation scale with sunlight and water. In the past, the Rubber Research Institute of Malaysia has generated transgenic Hevea that expressed recombinant human serum albumin, scFv antibody in latex serum. However, the production is still below the 1.0 % level for commercial feasibility. This study evaluates the promoter activity of hevein gene, which encodes the most abundant soluble protein in Hevea latex. Genetic constructs were prepared with the test hevein promoter fragments Hev P1 (0.35kb), Hev P2 (0.45kb) and Hev P3 (0.73kb) that were inserted 5’ to the reporter gene uidA in pGPTV-KAN expression vector. The expression vectors containing the hevein promoter fused to uidA in Agrobacterium tumefaciens GV2260 were then used to infect Hevea anther callus. Putative transgenic rubber plantlets generated via somatic embryogenesis tested for gene insertion, gave signals for single and mutiple copies for uidA gene and this was similarly reflected in the positive control (plants with 35S CaMV promoter) by Southern analysis. The recombinat protein levels for β-glucuronidase (GUS) in the latex of Hevea transformants and controls was quantified using ELISA technique. The levels of GUS in latex serum from positive controls (B6 plants) ranged from 0.063 to 0.079 µg/ml of latex serum. The levels of GUS protein (µg/ml of latex serum) in Hevea transformants P1 ranged from 0.0052 to 1.55; P2 ranged from 0.005 to 0.0012; P3 ranged from 0.0017 to 0.0027. The highest expressing transgenic plants was derived from B6 using the 35S CaMV promoter for uidA gene and P1 containing the smallest fragment of the hevein promoter for uidA gene. This study paves the way to sucessfully produce recombinant proteins of commercial value in transgenic rubber trees. 2011-06-14 Conference or Workshop Item NonPeerReviewed application/pdf en http://irep.iium.edu.my/2759/1/Arokiaraj_Pappusamy.22nd_Science_Congress_Conference_paper.2011.docx Pappusamy, Arokiaraj / PA and Shuib, Siti Shuhada and Enamalai, Sunderasan (2011) Functional analysis of a latex-specific promoter (hevein) of Hevea for enhanced recombinant protein production in Hevea brasiliensis. In: 22nd Pacific Science Congress, 14-17 June 2011, Kuala Lumpur Convention Centre, Malaysia.
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English
topic	SB Plant culture
spellingShingle	SB Plant culture Pappusamy, Arokiaraj / PA Shuib, Siti Shuhada Enamalai, Sunderasan Functional analysis of a latex-specific promoter (hevein) of Hevea for enhanced recombinant protein production in Hevea brasiliensis
description	Any protein, either naturally occurring or designed synthetically, could be produced safely, inexpensively and in large quantities using the power of the rubber trees for the production of recombinant proteins on a plantation scale with sunlight and water. In the past, the Rubber Research Institute of Malaysia has generated transgenic Hevea that expressed recombinant human serum albumin, scFv antibody in latex serum. However, the production is still below the 1.0 % level for commercial feasibility. This study evaluates the promoter activity of hevein gene, which encodes the most abundant soluble protein in Hevea latex. Genetic constructs were prepared with the test hevein promoter fragments Hev P1 (0.35kb), Hev P2 (0.45kb) and Hev P3 (0.73kb) that were inserted 5’ to the reporter gene uidA in pGPTV-KAN expression vector. The expression vectors containing the hevein promoter fused to uidA in Agrobacterium tumefaciens GV2260 were then used to infect Hevea anther callus. Putative transgenic rubber plantlets generated via somatic embryogenesis tested for gene insertion, gave signals for single and mutiple copies for uidA gene and this was similarly reflected in the positive control (plants with 35S CaMV promoter) by Southern analysis. The recombinat protein levels for β-glucuronidase (GUS) in the latex of Hevea transformants and controls was quantified using ELISA technique. The levels of GUS in latex serum from positive controls (B6 plants) ranged from 0.063 to 0.079 µg/ml of latex serum. The levels of GUS protein (µg/ml of latex serum) in Hevea transformants P1 ranged from 0.0052 to 1.55; P2 ranged from 0.005 to 0.0012; P3 ranged from 0.0017 to 0.0027. The highest expressing transgenic plants was derived from B6 using the 35S CaMV promoter for uidA gene and P1 containing the smallest fragment of the hevein promoter for uidA gene. This study paves the way to sucessfully produce recombinant proteins of commercial value in transgenic rubber trees.
format	Conference or Workshop Item
author	Pappusamy, Arokiaraj / PA Shuib, Siti Shuhada Enamalai, Sunderasan
author_facet	Pappusamy, Arokiaraj / PA Shuib, Siti Shuhada Enamalai, Sunderasan
author_sort	Pappusamy, Arokiaraj / PA
title	Functional analysis of a latex-specific promoter (hevein) of Hevea for enhanced recombinant protein production in Hevea brasiliensis
title_short	Functional analysis of a latex-specific promoter (hevein) of Hevea for enhanced recombinant protein production in Hevea brasiliensis
title_full	Functional analysis of a latex-specific promoter (hevein) of Hevea for enhanced recombinant protein production in Hevea brasiliensis
title_fullStr	Functional analysis of a latex-specific promoter (hevein) of Hevea for enhanced recombinant protein production in Hevea brasiliensis
title_full_unstemmed	Functional analysis of a latex-specific promoter (hevein) of Hevea for enhanced recombinant protein production in Hevea brasiliensis
title_sort	functional analysis of a latex-specific promoter (hevein) of hevea for enhanced recombinant protein production in hevea brasiliensis
publishDate	2011
url	http://irep.iium.edu.my/2759/ http://irep.iium.edu.my/2759/1/Arokiaraj_Pappusamy.22nd_Science_Congress_Conference_paper.2011.docx
first_indexed	2023-09-18T20:10:23Z
last_indexed	2023-09-18T20:10:23Z
_version_	1777407437707411456

Functional analysis of a latex-specific promoter (hevein) of Hevea for enhanced recombinant protein production in Hevea brasiliensis

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