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Cloning, purification, crystallization and X-ray crystallographic analysis of the periplasmic sensing domain of Pseudomonas fluorescens chemotactic transducer of amino acids type A (CtaA)

Biosci Trends.. 2016-09; 
Ud-Din AI, Roujeinikova A.
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Gene Synthesis ...The sequence for the periplasmic sensing domain (CtaAperi, residues 34-277) was codon-optimized for expression in Escherichia coli and synthesized and ligated into the expression vector pET151/D-TOPO (Invitrogen) by GenScript... Get A Quote

摘要

Chemotaxis towards nutrients plays a crucial role in root colonization by Pseudomonas fluorescens. The P. fluorescens chemotactic transducer of amino acids type A (CtaA) mediates movement towards amino acids present in root exudates. In this study, the periplasmic sensory domain of CtaA has been crystallized by the hanging-drop vapor diffusion method using ammonium sulfate as a precipitating agent. A complete data set was collected to 1.9 Å resolution using cryocooling conditions and synchrotron radiation. The crystals belong to space group I222 or I212121, with unit-cell parameters a = 67.2, b = 76.0, c = 113.3 Å. This is an important step towards elucidation of the structural basis of how CtaA recognizes it... More

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