Comparison of renografin density gradient centrifugation and ion-exchange chromatography for purification of phony peach bacterium from plant extracts |
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Authors: | William E. Kuriger Norman W. Schaad William J. French |
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Affiliation: | (1) Georgia Experiment Station, University of Georgia, 30212 Experiment, Georgia, USA;(2) Institute of Food and Agricultural Science, Agriculture Research Center, University of Florida, 32344 Monticello, Florida, USA |
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Abstract: | Renografin density gradient centrifugation and carboxymethyl (CM)-and diethylaminoethyl (DEAE)-cellulose ion-exchange chromatography were compared for purification of cells of phony peach, rickettsia-like bacterium (RLB) from plant extracts. Centrifugation on 30–40% linear gradients resulted in four bands. Cells of RLB sedimented into the third band located 4.9–5.4 cm below the meniscus. Phase-contrast microscopy, plant peroxidase activity, and viable bacterial assays showed the resulting cells ofRLB to be relatively free of host contaminants. A typical purification from 120 ml of extract containing 7.2×109RLB cells resulted in the recovery of 7.0×107RLB cells. Cells ofRLB were not successfully purified byCM andDEAE chromatography. Unitil a medium is available for cultivation of plantRLB, Renografin density gradient centrifugation should be a useful tool for obtaining cells ofRLB for biochemical and serological investigations. |
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