کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
36013 45118 2007 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Isolation and mutation of recombinant EPSP synthase from pathogenic bacteria Pseudomonas aeruginosa
موضوعات مرتبط
مهندسی و علوم پایه مهندسی شیمی بیو مهندسی (مهندسی زیستی)
پیش نمایش صفحه اول مقاله
Isolation and mutation of recombinant EPSP synthase from pathogenic bacteria Pseudomonas aeruginosa
چکیده انگلیسی

The Pseudomonas aeruginosa aroA gene encodes an enzyme called 5-enol-pyruvylshikimate-3-phosphate (EPSP) synthase, which has been shown as the primary target of the herbicide glyphosate. We have cloned this gene and constructed a system for the high level expression of a recombinant form of this enzyme by amplifying the aroA gene from the P. aeruginosa genomic DNA and subcloning into a vector suitable for expression in Escherichia coli. The resulting plasmid, pTrcPA, produced the EPSP synthase in large quantities which has been purified to homogeneity. Furthermore, the site-directed mutants of P. aeruginosa ESPS synthase have been constructed in order to compare in vitro glyphosate sensitivity between the wild-type and the mutant enzymes. The kcat and Km values for substrates in both forward and reverse reactions were obtained from both wild-type and mutant EPSP synthases.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Process Biochemistry - Volume 42, Issue 4, April 2007, Pages 592–598
نویسندگان
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