Isochorismate pyruvate lyase: Difference between revisions
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==Applications== | ==Applications== | ||
Bacterial genes can be introduced into plants to increase the accumulation of the desired compounds. Salicylic acid content in ''Arabidopsis thaliana'' can be manipulated by expressing an engineered bacterial salicylate synthase by the fusion of two bacterial genes pchA and pchB from the human pathogen, ''Pseudomonas aeruginosa'', that encode isochorismate synthase and isochorismate pyruvate lyase. These are expressed under a constitutive promoter. | Bacterial genes can be introduced into plants to increase the accumulation of the desired compounds. Salicylic acid content in ''Arabidopsis thaliana'' can be manipulated by expressing an engineered bacterial salicylate synthase by the fusion of two bacterial genes pchA and pchB from the human pathogen, ''Pseudomonas aeruginosa'', that encode isochorismate synthase and isochorismate pyruvate lyase. These are expressed under a constitutive promoter. | ||
==Crystallization== | |||
X-ray crystallization is performed by hanging drop method for the apo, open and closed structures. The apo structure has nitrate in it in the regions that have the pyruvate in the bound structures. In the case of the bound structures, immediately prior to crystallization dilution in pyruvate is performed. | |||
These proteins carry the 6His Tag, but on crystallization, these are proteolyzed in the drop. | |||
Molecular replacement of the Apo with the structure of ECM (chorismate mutase) was done. Crystals of the bound structure are obtained by growing in PEG 3350. | |||