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However, the protective efficacy against VHSV challenge conferred by both plasmids was similar.
In the course of our studies on gene expression regulated by several GATA transcription factors, we employed both pRL-TK and pRL-SV40 as control reporter plasmids and observed that Rluc expression by both plasmids was increased by members of the GATA transcription factor family.
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(D ) Growth by ∆ysp2 cells expressing Ysp2p or the G > R mutant under the YSP2 promoter ± AmB. ‡ indicates that Ysp2S2 in both plasmids was substituted by Lam4S2 (53% identical, 72% homologous).
Both plasmids were verified by sequencing.
Both plasmids were transferred by conjugation to the derivative of S. Gallinarum strain 287/91 carrying SPI-19 flanked by loxP sites.
After 2 h incubation at 37°C, cells containing both plasmids were selected by using a 96-prong frogger to transfer cells to LB plates containing ampicillin and tetracycline.
Both plasmids were transferred by conjugation, belong to the incompatibility group F and represent two replicons of type FII and FIA (Table S3 and S4, Fig. 3).
Both plasmids were transfected by using FuGene6 (Roche, Penzberg, Germany) according to manufacturer's protocol.
This has previously been reported, and appears to be an artifact of the technologies used, as both plasmids are detectable by PCR [ 40].
The cDNAs in both plasmids were driven by a strong human cytomegalovirus immediate early promoter and each contained an intron to promote high-level expression.
CaM was cloned into the co-expression compatible plasmid pOKD454 and both plasmids were co-transformed by electroporation in BL21 DE3) cells (Novagen).
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