pMTL-YN1，pMTL-YN2，pMTL-YN1C， pMTLYN2C艰难梭菌基因敲入质粒Clostridioides difficile Knock-In plasmid BioVector NTCC Inc.

pMTL-YN1，pMTL-YN2，pMTL-YN1C， pMTLYN2C艰难梭菌基因敲入质粒Clostridioides difficile Knock-In plasmid vector

BioVector NTCC Inc.

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A series of pyrE-basedACE vectors are available equivalent to those made for C. acetobutylicum.  All carry the C. perfringenscatP gene conferring thiamphenicol resistance; the replication region of the E.coli plasmid ColE1, and the TraJ transfer function of the RP4 oriT region. Plasmid pMTL-YN1, and derivatives, carry the pCB102 Gram +ve replicon and is intended for use in the 630Δerm strain, whereas plasmid pMTL-YN2, and derivatives, carries the pBP1 Gm +ve replicon and is intended for use in RT027 strains, such as R20291 [1].

Correction Vectors pMTL-YN1 and pMTLYN2
Used to restore the mutant pyrE locus to wildtype – intended for use when the phenotype of mutations made elsewhere in the genome, using the KO vectors pMTL-YN3 and pMTL-YN4, needs to be assessed. Equivalent to the C. acetobutylicum plasmid pMTL-ME6 [2].

Complementation Vectors pMTL-YN1C and pMTLYN2C
Used to deliver a functional copy of a knocked-out gene (using pMTL-YN3 and pMTL-YN4) into the genome at the pyrE locus, while at the same time restoring the cell to prototrophy. Compared to pMTL-YN1 and pMTL-YN2,  plasmids pMTL-YN1C and pMTL-YN2C have an additional segment of DNA inserted between the left-hand homology arm (LHA) and the right-hand homology arm (RHA) which carries: a transcriptional terminator (T1) of the ferredoxin gene of Clostridium pasteurianum; a copy of the lacZ’ gene encoding the alpha fragment of the E.coli β-galactosidase, and; a multiple cloning site (MCS) region derived from plasmid pMTL20

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