Nucleic Acid Probes for Detection of Phytopathogenic Bacteria. Phase 1 Final Report

摘要

The study examined the feasibility of developing a diagnostic procedure for bacterial diseases in crop plants based on nucleic acid hybridization technologies. Erwinia carotovora var. atroseptica (ECa) was chosen as the test organism for the study. A library of ECa clones was prepared by cloning into the PUC8 plasmid and transforming C600. The ECa inserts ranged in size from 400-1200 base pairs. These clones were screened with RTase labelled ECa DNA and E. herbicola DNA by colony hybridization. The hybridization patterns were then compared. A two-dimensional gel system was also used to discern sequence mismatches between S1 digested heteroduplexes of ECa and E. herbicola DNA digested with a four-base pair recognition restriction enzyme.