Stjepandic, D., Weinel, C., Hilbert, H., Koo, H. L., Diehl, F., Nelson, K. E., Tummler, B., Hoheisel, J. D.
The Genome Structure of Pseudomonas putida : High-resolution Mapping and Microarray Analysis
Environ Microbiol. 2002 Dec 01; 4(12): 819-23.
As part of a collaborative project aimed at sequencing and functionally analysing the entire genome of Pseudomonas putida strain KT2440, a physical clone map was produced as an initial resource. To this end, a high-coverage cosmid library was arrayed and ordered by clone hybridizations. Restriction fragments generated by rare-cutting enzymes and plasmids containing the rrn operon and 23S rDNA of Pseudomonas aeruginosa were used as probes and, parts of the cosmids were end-sequenced. This provided the information necessary for merging and comparing the macro-restriction map, cosmid clone order and sequence information, thereby assuring co-linearity of the eventual sequence assembly with the actual genome. A tiling path of clones was selected, from the shotgun clones used for sequencing, for the production of DNA microarrays that represent the entire genome including its non-coding portions.