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A Thermostable beta-Glucuronidase Obtained by Directed Evolution as a Reporter Gene in Transgenic Plants

文献类型： 外文期刊

作者： Xiong, Ai-Sheng 1 ; Peng, Ri-He 2 ; Zhuang, Jing 3 ; Chen, Jian-Min 4 ; Zhang, Bin 2 ; Zhang, Jian 3 ; Yao, Quan-Hong 2 ;

作者机构： 1.Nanjing Agr Univ, State Key Lab Crop Genet & Germplasm Enhancement, Coll Hort, Nanjing, Peoples R China

2.Shanghai Acad Agr Sci, Biotechnol Res Inst, Shanghai, Peoples R China

3.Alberta Innovates Technol Futures, Vegreville, AB, Canada

4.Yangzhou Univ, Coll Biosci & Biotechnol, Yangzhou, Peoples R China

期刊名称：PLOS ONE （ 影响因子：3.24； 五年影响因子：3.788 ）

ISSN： 1932-6203

年卷期： 2011 年 6 卷 11 期

页码：

收录情况： SCI

摘要： A beta-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 b-glucuronidase activity after exposure to 60 degrees C for 10 min, while those hosting the gus-tr3337 gene retained 70% or 50% activity after exposure to 80 degrees C for 10 min or 30 min, respectively. Similarly, in vivo b-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 degrees C for up to 30 min. Thus, the thermostability of GUS-TR3337 can be exploited to distinguish between endogenous and transgenic beta-glucuronidase activity, which is a welcome improvement in its use as a reporter.

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