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The construct was transformed into tpx mutant by electroporation and gentamicin resistant transformants were selected.
After transforming the pCB1532::MPG1 into ΔpdeH mutant, the SUR resistant transformants were screened by qRT-PCR with primers MPG1_QF/MPG1_QR.
Basta resistant transformants were selected under greenhouse conditions.
Chloramphenicol (15 µg/ml) resistant transformants were then selected.
After 1 to 2 weeks of incubation, spectinomycin resistant transformants were used to inoculate liquid medium.
Southern blot analysis of ten tetracycline resistant transformants demonstrated the intended deletion in the mg192 gene (Fig. 8C).
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During development of the transformation protocol, transformation parameters were modified until paromomycin-resistant transformants were obtained (see below).
The resulting cassette was integrated into K12 by lithium acetate transformation and stable G418-resistant transformants were selected.
A total of ∼280 erythromycin (Erm -resistant transformants wErm -resistantn each of transformantsdent transformation experiments were strain 13.
The construct was transformed into the gi-2 mutant background and kanamycin-resistant transformants selected.
In the studies described above, the PeUMPS cDNA was introduced into strain M4 using a co-transformation technique with pG418-T1A plasmid to first select G418-resistant transformants.
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