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Concurrent mutations in three amino acids in 5-enopyruvylshikimate-3-phosphate synthase from Ochrobactrum improve glyphosate resistance.
MedLine Citation:
PMID:  21948846     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
A mutant of 5-enopyruvylshikimate-3-phosphate synthase from Ochrobactrum anthropi has been identified after four rounds of DNA shuffling and screening. Its ability to restore the growth of the mutant ER2799 cell on an M9 minimal medium containing 300 mM glyphosate led to its identification. The mutant had mutations in seven amino acids: E145G, N163H, N267S, P318R, M377V, M425T, and P438L. Among these mutations, N267S, P318R, and M425T have never been previously reported as important residues for glyphosate resistance. However, they were found to collectively contribute to the improvement of glyphosate tolerance using site-directed mutagenesis in the present study. Kinetic analyses of these three mutants demonstrated that the effectiveness of these three individual amino acids alterations on glyphosate tolerance was in the order P318R > M425T > N267S. The results of the kinetic analyses combined with a three-dimensional structure modeling of the location of P318R and M425T demonstrate that the down-hemisphere surface is another important region for glyphosate resistance. Furthermore, the transgenic Arabidopsis was obtained to confirm the potential of the mutant in developing glyphosate-resistant crops.
Authors:
Yong-Sheng Tian; Jing Xu; Ai-Sheng Xiong; Wei Zhao; Xiao-Yan Fu; Ri-He Peng; Quan-Hong Yao
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Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2011-9-23
Journal Detail:
Title:  Applied and environmental microbiology     Volume:  -     ISSN:  1098-5336     ISO Abbreviation:  -     Publication Date:  2011 Sep 
Date Detail:
Created Date:  2011-9-27     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7605801     Medline TA:  Appl Environ Microbiol     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Affiliation:
Shanghai Key Laboratory of Agricultural Genetics and Breeding, Biotechnology Research Institute, Shanghai Academy of Agricultural Sciences, 2901 Beidi Road, Shanghai, People's Republic of China.
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