For a given insertion site, different inserted sequences give rise to different stabilities, but the relationship between stability and sequence is not yet clear.
A BLASTX analysis [ 20] of the inserted sequences allowed us to identify a single LAGLIDADG ORF in each insertion.
The inserted sequences were regulated by a spore-specific promoter located upstream of p19.
Approximately 1.5 Mb of inserted sequences were identified, including 743 kb containing 615 ORFs that are absent from published P. aeruginosa genomes.
The inserts caused no adverse effects on viral infectivity and virulence, and the inserted sequences remained intact in progeny viruses in the systemically infected leaves.
In the recombinant B. subtilis strains, the inserted sequences were located downstream of a strong phage promoter that could be activated by thermal induction.
Using a plasmid vector system to survey different inserted sequences for their dependence on introns for expression, we found that the presence of IVS2 stimulated the expression of these sequences 2- to 500-fold.
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Colonies containing plasmids with inserted sequences were initially screened by DNA dot hybridization, and those positive colonies were then screened by hybrid selected translation.
The visible DNA bands on the gels of sample 1, sample 2 and sample 3 were sequenced, and the chromatograms showed that these band all contained target sequences adjacent to the inserted sequences (Figs. 2b d, 3b d, 4b d).
All the inserted sequences were confirmed by sequencing.
