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We have demonstrated in this report that the construction of libraries for the E. coli and B. subtilis genomes without passaging through E. coli is practical, but the number of transformants is less than for libraries cloned using E. coli as a host.
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Proliferation of Bax/Bcl-w transformants was less dramatically impaired.
However, the decrease in ethidium bromide accumulation in the transformants was less pronounced than that in M184, which accumulated 2.6-fold lower levels of ethidium bromide than R6.
Transformation of ArntbHLH and Arnt-C/EBP was repeated, and transformants were plated under less stringent conditions (5 mM 3-AT) to ensure that the previous results were not false negatives.
Because the Rmontanensis700 transformant produced unexpected GFPuv mRNA transcripts and it was less fluorescent than the Rmona658 transformant that produced the expected GFPuv mRNA transcript [39], we used Western immunoblots to study expression of the GFPuv protein.
Kanr or Camr transformants of strain M1.5 made with these constructs were less Mtz resistant (phenotype 160R,190S) than were those containing the fur-wt allele (190R,220S) (Fig. 4A).
Those transformants that had received the fur-wt allele had a 190R,220S phenotype, and were less Mtz resistant than isogenic Camr siblings that had retained furR3I (230R,250S).
In contrast, only a small proportion of transformants were coloured when the reporter was expressed from Phsp60; less than 1% of the transformants for tdTomato, Turbo and 5% for mCherry.
The transformants were cultured and selected.
Transformants were selected on SD-URA plates.
One of the Ura+ transformants was named strain TR2-7.
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