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通过十八烷基硅烷固相吸附从番茄链格孢菌中快速纯化宿主特异性致病毒素。

Rapid purification of host-specific pathotoxins from Alternaria alternata f. sp. lycopersici by solid-phase adsorption on octadecylsilane.

作者信息

Clouse S D, Martensen A N, Gilchrist D G

出版信息

J Chromatogr. 1985 Dec 20;350(1):255-63. doi: 10.1016/s0021-9673(01)93524-1.

Abstract

A simplified, rapid procedure for the purification of two phytotoxic metabolites (TA and TB) from cell-free culture filtrates of Alternaria alternata f. sp. lycopersici was developed using hydrophobic chromatography on macroporous, C18 bonded silica followed by gel filtration. Baseline separation of TA from TB was achieved in the final preparation and yields of 80% were obtained with retention of full biological activity in all assay systems tested. Toxin analysis by nuclear magnetic resonance and high-performance liquid chromatography indicated TA was at least 99% pure. The efficiency of the procedure allows rapid accumulation of pure TA and TB for studies on the molecular mode of action and host plant response to this novel host-specific phytotoxin.

摘要

开发了一种简化、快速的方法,用于从番茄链格孢菌的无细胞培养滤液中纯化两种具有植物毒性的代谢物(TA和TB)。该方法先采用大孔C18键合硅胶进行疏水色谱,然后进行凝胶过滤。最终制备物中实现了TA与TB的基线分离,产率达到80%,且在所有测试的分析系统中均保留了全部生物活性。通过核磁共振和高效液相色谱进行的毒素分析表明,TA的纯度至少为99%。该方法的高效性使得能够快速积累纯TA和TB,用于研究这种新型寄主特异性植物毒素的分子作用模式和寄主植物反应。

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