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REPEATED SEQUENCES INCLUDING RS1100 FROM PSEUDOMONAS CEPACIA AC100 FUNCTION AS IS ELEMENTS
Citation:
Haughland, R., U. Sangodkar, AND A. Chakrabarty. REPEATED SEQUENCES INCLUDING RS1100 FROM PSEUDOMONAS CEPACIA AC100 FUNCTION AS IS ELEMENTS. U.S. Environmental Protection Agency, Washington, D.C., EPA/600/J-93/235.
Description:
Several lines of evidence were obtained that the previously identified, repeated sequence RS1100 of Pseudomonas cepacia strain AC1100 undergoes transposition events. NA sequences flanking the chlorohydroxy hydroquinone (CHQ) degradative genes of this organism were examined from sources, including several independently isolated cosmid clones from an AC1100 genomic library and genomic DNAs of two independently maintained wild-type AC1100 isolates. ybridization and restriction endonuclease mapping studies revealed these sequences to be similar except for their numbers and distributions of RS1100 copies. ecombinant plasmid containing the immediate chq gene region and excluding any copies of RS1100 was conjugated into AC1100 mutant RHA5 which was shown to have undergone a deletion of this corresponding DNA. ybridization and restriction mapping analyses of several reisolated plasmids revealed the presence of RS1100 sequences at different positions within either the vector or insert portions. ne such plasmid contained tandem copies of RS1100 with an intervening DNA sequence also of AC1100 origin. imilar experiments involving introduction of the promoter probe plasmid pKT240 into wild-type AC1100 cells resulted in the acquisition of high-concentration streptomycin resistance by a number of recipients. he reisolated plasmids in most cases also conferred streptomycin resistance to Escherichia oli ransformants nd n each case were found to contain insertions close to the upstream portion of the aphC structural gene. hese insertions alternatively contained RS1100 sequences or a newly identified 3400 bp repeated sequence from AC1100.