A thermostable β-glucuronidase obtained by directed evolution as a reporter gene in transgenic plants.

A β-glucuronidase variant, GUS-TR3337, that was obtained by directed evolution exhibited higher thermostability than the wild-type enzyme, GUS-WT. In this study, the utility of GUS-TR337 as an improved reporter was evaluated. The corresponding gus-tr3337 and gus-wt genes were independently cloned in...

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Autores principales:	Ai-Sheng Xiong, Ri-He Peng, Jing Zhuang, Jian-Min Chen, Bin Zhang, Jian Zhang, Quan-Hong Yao
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/c853f908cfa04ba4899669ba9b3c636b
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spelling	oai:doaj.org-article:c853f908cfa04ba4899669ba9b3c636b2021-11-18T07:34:41ZA thermostable β-glucuronidase obtained by directed evolution as a reporter gene in transgenic plants.1932-620310.1371/journal.pone.0026773https://doaj.org/article/c853f908cfa04ba4899669ba9b3c636b2011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22096498/?tool=EBIhttps://doaj.org/toc/1932-6203A β-glucuronidase variant, GUS-TR3337, that was obtained by directed evolution exhibited higher thermostability than the wild-type enzyme, GUS-WT. In this study, the utility of GUS-TR337 as an improved reporter was evaluated. The corresponding gus-tr3337 and gus-wt genes were independently cloned in a plant expression vector and introduced into Arabidopsis thaliana. With 4-MUG as a substrate, plants containing the gus-wt gene showed no detectable β-glucuronidase activity after exposure to 60°C for 10 min, while those hosting the gus-tr3337 gene retained 70% or 50% activity after exposure to 80°C for 10 min or 30 min, respectively. Similarly, in vivo β-glucuronidase activity could be demonstrated by using X-GLUC as a substrate in transgenic Arabidopsis plants hosting the gus-tr3337 gene that were exposed to 80°C for up to 30 min. Thus, the thermostability of GUS-TR3337 can be exploited to distinguish between endogenous and transgenic β-glucuronidase activity, which is a welcome improvement in its use as a reporter.Ai-Sheng XiongRi-He PengJing ZhuangJian-Min ChenBin ZhangJian ZhangQuan-Hong YaoPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 11, p e26773 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ai-Sheng Xiong Ri-He Peng Jing Zhuang Jian-Min Chen Bin Zhang Jian Zhang Quan-Hong Yao A thermostable β-glucuronidase obtained by directed evolution as a reporter gene in transgenic plants.
description	A β-glucuronidase variant, GUS-TR3337, that was obtained by directed evolution exhibited higher thermostability than the wild-type enzyme, GUS-WT. In this study, the utility of GUS-TR337 as an improved reporter was evaluated. The corresponding gus-tr3337 and gus-wt genes were independently cloned in a plant expression vector and introduced into Arabidopsis thaliana. With 4-MUG as a substrate, plants containing the gus-wt gene showed no detectable β-glucuronidase activity after exposure to 60°C for 10 min, while those hosting the gus-tr3337 gene retained 70% or 50% activity after exposure to 80°C for 10 min or 30 min, respectively. Similarly, in vivo β-glucuronidase activity could be demonstrated by using X-GLUC as a substrate in transgenic Arabidopsis plants hosting the gus-tr3337 gene that were exposed to 80°C for up to 30 min. Thus, the thermostability of GUS-TR3337 can be exploited to distinguish between endogenous and transgenic β-glucuronidase activity, which is a welcome improvement in its use as a reporter.
format	article
author	Ai-Sheng Xiong Ri-He Peng Jing Zhuang Jian-Min Chen Bin Zhang Jian Zhang Quan-Hong Yao
author_facet	Ai-Sheng Xiong Ri-He Peng Jing Zhuang Jian-Min Chen Bin Zhang Jian Zhang Quan-Hong Yao
author_sort	Ai-Sheng Xiong
title	A thermostable β-glucuronidase obtained by directed evolution as a reporter gene in transgenic plants.
title_short	A thermostable β-glucuronidase obtained by directed evolution as a reporter gene in transgenic plants.
title_full	A thermostable β-glucuronidase obtained by directed evolution as a reporter gene in transgenic plants.
title_fullStr	A thermostable β-glucuronidase obtained by directed evolution as a reporter gene in transgenic plants.
title_full_unstemmed	A thermostable β-glucuronidase obtained by directed evolution as a reporter gene in transgenic plants.
title_sort	thermostable β-glucuronidase obtained by directed evolution as a reporter gene in transgenic plants.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/c853f908cfa04ba4899669ba9b3c636b
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A thermostable β-glucuronidase obtained by directed evolution as a reporter gene in transgenic plants.

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