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Source of tetracycline promoter (PTet) (Ishiwa and Shibahara 1985) pQF50 A broad-host range vector.
All plasmids for overexpression of phaZ genes were derived from the broad host range vector pBBR1MCS-2 (Table 2).
A genetic construction was carried out using the broad host range vector pKT230 and plasmid pMCG898, which encodes the Erwinia herbicola pyrroloquinoline quinone (PQQ) synthase, a gene involved in mineral phosphate solubilization (mps).
We then cloned the pvrR gene into the broad host range vector pBBR1MCS-4, yielding pBBR1MCS-4-pvrR (Table 2), and introduced it into PA14::rcsC::cupD1-lacZ.
Furthermore, the rcsB gene was cloned under the control of the lac promoter into the broad host range vector pBBR1MCS-5 and introduced into PA14::cupD1-lacZ.
Overexpression constructs were obtained by PCR amplifying the gene of interest, cloning into pCR2.1-TA and sub-cloning into a pBBR broad host range vector.
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To address these problems, we have designed and constructed a set of broad host range vectors that are transferable via intergeneric conjugation with an Escherichia coli strain carrying chromosomally-encoded transfer functions.
As was pointed out by Comaniciu and Meer (2002), when the location and range vectors are concatenated in the joint spatial-range domain of dimension d = p + 2, their different natures have to be compensated by proper normalization of parameters h s and h r.
The slant-range vector from the aft antenna phase center to the target can, therefore, be expressed as. (36a).
As discussed in Materials and Methods, the broad host-range vector pBBR1MCS-2 was used as a parental plasmid to construct the phaZ overexpression plasmids.
Plasmid pRK404 a smaller derivative of RK2 is a tetracycline-resistant broad-host-range vector that carries a multiple cloning site and the lacZ peptide that enables blue/white selection for cloned inserts in Escherichia coli.
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