Purification of the native enzyme and cloning and characterization of a cDNA for (+)-delta-cadinene synthase from bacteria-inoculated cotton foliar tissue.

摘要

Scope and method of study. The first objective of this study was to purify (+)-;Findings and conclusions. CDN1 was purified to apparent electrophoretic homogeneity using a combination of salt-induced phase separation, batch-mode hydroxylapatite fractionation, hydrophobic-interaction and strong anion-exchange chromatography, and renaturation following denaturing polyacrylamide gel electrophoresis. Amino acid sequences for three tryptic peptides were determined and used in the cloning of two isozymes, one of which, when overexpressed in E. coli demonstrated CDN1 activity. Preliminary transcript expression studies using plants inoculated in the cotyledons with Xcm, showed the induction of both cdn1s to peak at ca. 24 hours post inoculation (hpi) in the cotyledons, while a systemic induction of one cdn1 occurred at ca. 36 hpi in the roots.