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Title: Improved Plant-based Production of E1 endoglucanase Using Potato: Expression Optimization and Tissue Targeting

Abstract

Optimization of Acidothermus cellulolyticus endoglucanase (E1) gene expression in transgenic potato (Solanum tuberosum L.) was examined in this study, where the E1 coding sequence was transcribed under control of a leaf specific promoter (tomato RbcS-3C) or the Mac promoter (a hybrid promoter of mannopine synthase promoter and cauliflower mosaic virus 35S promoter enhancer region). Average E1 activity in leaf extracts of potato transformants, in which E1 protein was targeted by a chloroplast signal peptide and an apoplast signal peptide were much higher than those by an E1 native signal peptide and a vacuole signal peptide. E1 protein accumulated up to 2.6% of total leaf soluble protein, where E1 gene was under control of the RbcS-3C promoter, alfalfa mosaic virus 5-untranslated leader, and RbcS-2A signal peptide. E1 protein production, based on average E1 activity and E1 protein accumulation in leaf extracts, is higher in potato than those measured previously in transgenic tobacco bearing the same transgene constructs. Comparisons of E1 activity, protein accumulation, and relative mRNA levels showed that E1 expression under control of tomato RbcS-3C promoter was specifically localized in leaf tissues, while E1 gene was expressed in both leaf and tuber tissues under control of Mac promoter. This suggestsmore » dual-crop applications in which potato vines serve as enzyme production `bioreactors' while tubers are preserved for culinary applications.« less

Authors:
 [1];  [1];  [1];  [2]
  1. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
  2. National Renewable Energy Lab. (NREL), Golden, CO (United States)
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
15001181
Report Number(s):
PNNL-SA-32097
Journal ID: ISSN 1380-3743; 820101000
DOE Contract Number:  
AC06-76RL01830
Resource Type:
Journal Article
Journal Name:
Molecular Breeding
Additional Journal Information:
Journal Volume: 6; Journal Issue: 3; Journal ID: ISSN 1380-3743
Publisher:
Springer
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Dai, Ziyu, Hooker, Brian S., Anderson, Daniel B., and Thomas, Steven R. Improved Plant-based Production of E1 endoglucanase Using Potato: Expression Optimization and Tissue Targeting. United States: N. p., 2000. Web. doi:10.1023/A:1009653011948.
Dai, Ziyu, Hooker, Brian S., Anderson, Daniel B., & Thomas, Steven R. Improved Plant-based Production of E1 endoglucanase Using Potato: Expression Optimization and Tissue Targeting. United States. doi:10.1023/A:1009653011948.
Dai, Ziyu, Hooker, Brian S., Anderson, Daniel B., and Thomas, Steven R. Thu . "Improved Plant-based Production of E1 endoglucanase Using Potato: Expression Optimization and Tissue Targeting". United States. doi:10.1023/A:1009653011948.
@article{osti_15001181,
title = {Improved Plant-based Production of E1 endoglucanase Using Potato: Expression Optimization and Tissue Targeting},
author = {Dai, Ziyu and Hooker, Brian S. and Anderson, Daniel B. and Thomas, Steven R.},
abstractNote = {Optimization of Acidothermus cellulolyticus endoglucanase (E1) gene expression in transgenic potato (Solanum tuberosum L.) was examined in this study, where the E1 coding sequence was transcribed under control of a leaf specific promoter (tomato RbcS-3C) or the Mac promoter (a hybrid promoter of mannopine synthase promoter and cauliflower mosaic virus 35S promoter enhancer region). Average E1 activity in leaf extracts of potato transformants, in which E1 protein was targeted by a chloroplast signal peptide and an apoplast signal peptide were much higher than those by an E1 native signal peptide and a vacuole signal peptide. E1 protein accumulated up to 2.6% of total leaf soluble protein, where E1 gene was under control of the RbcS-3C promoter, alfalfa mosaic virus 5-untranslated leader, and RbcS-2A signal peptide. E1 protein production, based on average E1 activity and E1 protein accumulation in leaf extracts, is higher in potato than those measured previously in transgenic tobacco bearing the same transgene constructs. Comparisons of E1 activity, protein accumulation, and relative mRNA levels showed that E1 expression under control of tomato RbcS-3C promoter was specifically localized in leaf tissues, while E1 gene was expressed in both leaf and tuber tissues under control of Mac promoter. This suggests dual-crop applications in which potato vines serve as enzyme production `bioreactors' while tubers are preserved for culinary applications.},
doi = {10.1023/A:1009653011948},
journal = {Molecular Breeding},
issn = {1380-3743},
number = 3,
volume = 6,
place = {United States},
year = {2000},
month = {6}
}