Expression of a bacterial aroA mutant, aroA-M1, encoding 5-enolpyruvylshikimate-3-phosphate synthase for the production of glyphosate-resistant tobacco plants.
Glyphosate is a non-selective broad-spectrum herbicide that inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), a key enzyme in the aromatic amino acid biosynthetic pathway in microorganisms and plants. We have previously reported a strategy for engineering glyphosate-resistant class I EPSPS based on staggered-PCR technology. Selected mutant enzymes exhibited high K(i)[glyphosate] and low K(m)[PEP] values compared to the parental enzymes from Escherichia coli ( EcaroA) and Salmonella typhimurium ( StaroA). One mutant, aroA-M1, was further engineered with a tobacco chloroplast leader sequence, and then placed in the binary vector pCAMBIA1300 for Agrobacterium-mediated gene transfer to tobacco ( Nicotiana tabacum cv. Xanthi). Transgenic plants with increased resistance to glyphosate were generated.
Wang
He-Yong
HY
Biotechnology Research Center, The Key Laboratory of Gene Engineering of the Education Ministry, Zhongshan University, 51027, Guangzhou, People's Republic of China.
Li
Yun-Feng
YF
Xie
Long-Xu
LX
Xu
Peilin
P
D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D019883:Alkyl and Aryl Transferases / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D017931:DNA Primers; D004351:Drug Resistance; D004926:Escherichia coli / Q000201:enzymology; D029968:Escherichia coli Proteins / Q000235:genetics; D005818:Genetic Engineering; D005998:Glycine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D006540:Herbicides / Q000494:pharmacology*; D016296:Mutagenesis; D030821:Plants, Genetically Modified / Q000201:enzymology*; D016133:Polymerase Chain Reaction / Q000379:methods; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012486:Salmonella typhimurium / Q000201:enzymology; D014026:Nicotiana / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*; D000097797:Glyphosate
D051229:3-Phosphoshikimate 1-Carboxyvinyltransferase; D019883:Alkyl and Aryl Transferases; D001426:Bacterial Proteins; D017931:DNA Primers; D029968:Escherichia coli Proteins; D005998:Glycine; D006540:Herbicides; D000097797:Glyphosate