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Table 6 Bacterial strains, plasmids and primers used in the construction of the malT mutant

From: malT knockout mutation invokes a stringent type gene-expression profile in Actinobacillus pleuropneumoniae in bronchoalveolar fluid

Bacterial strains, plasmids or primers

Characteristic or sequence

Source or Remark

E. coli DH5a

F-φ80lac ZΔM15Δ(lac ZYA-rg F)U169 deo R rec A1 end A1 hsd R17(rk -, mk +) sup E44 thi-1 gyr A96 rel A1 λ-

Clonetech

E. coli β2155

thr B1004pro thi hsdS lacZΔ M15 (F'lacZΔM15lacIq traD 36 proA+ proB+) Δdap::erm(Ermr)recA::RP4-2-tet(Tcr)Mu- km(Kmr)λpir

Reference no. 28

E. coli DH5α-pTOPOPCR-malT

DH5α harboring pCR4-TOPO containing malT of A. pleuropneumiae CM5

This work

E. coli DH5α- pTopoMC

DH5a harboring pCR4-TOPO containing ΔmalT::cat

This work

E. coli DH5α-pEMOC2M

DH5a harboring pEMOC2 containing ΔmalT::cat

This work

A. pleuropneumoniae

MalT negative mutant of A.

This work

CM5 3ΔmalT

pleuropneumonaie CM5

 

pCR4-TOPO

A linearized plasmid for cloning PCR product

Invitrogen

pEMOC2

A conjugation vector based on pBluesript SK with mob RP4 and Cmr

Reference no. 31

pTOPOPCR-malT

pCR4-TOPO containing malT of A. pleuropneumiae CM5

This work

pTopoMC

pCR4-TOPO containing ΔmalT::cat

This work

pEMOC2M

harboring pEMOC2 containing ΔmalT::cat

This work

malT-L

malT-R

ATGCAAGCAACATTTTCAAGA

TTAGCTATACCCCATCATTCTCAA

Primers for amplification of the malT gene of A

pleuropneumoniae CM5

stopupmalT-L

TTAGTTAGTTACGAGCTTTTTCACACCGTTT

Primers for generation of a linearized plasmid containing a deletion of 900 bp in its malT gene cloned in pTOPOPCR-malT.

stopupmalT-R

TAACTAACTAATGGGAATGGCATCATTTAGA

 

pnmalT-L

TCATCTGCAGATGCAAGCAACATTTTCAAGA

Primers for amplication of the ΔmalT::cat and the insertion of the Pst I and Not I sites into the PCR product.

pnmalT-R

ACAATACAGCGGCCGCTTAGCTATACCCCATCATTCTCAA

 

cat-L

CGGTGCCCTGAATGAACT

Primers for the PCR

cat-R

AAGCTTCGACGAATTTCTGC

amplification of omlA-P driven cat gene of pEMOC2