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Table 1 Characteristics of bacterial strains, plasmids, and primers used in this study

From: Attenuated Actinobacillus pleuropneumoniae double-deletion mutant S-8∆clpP/apxIIC confers protection against homologous or heterologous strain challenge

Strains, plasmids, and primers

Characteristics or sequence

Source or reference

Strains

E. coli β2155

thrB1004 pro thi strA hsdS lacZM15 (F’ lacZM15 lacl q traD36 proA + proB +)dap :: erm (Ermr))recA :: RPA-2-tet(Tcr)::Mu-km (Kmr) λpir

[21]

A.pleuropneumoniae S-8

A. pleuropneumoniae serotype 7 clinical isolate from the lung of a diseased pig in northern China

[17]

A.pleuropneumoniae S-8clpP

Unmarked clpP gene knockout mutant of A. pleuropneumoniae S-8

[17]

A.pleuropneumoniae S-8clpPapxIIC

Unmarked clpP/apxIIC genes knockout mutant of A. pleuropneumoniae S-8

This work

Plasmids

 pEMOC2

Conjugative vector based on pBluescript SK with mob RP4, polycloning site, Cm r, and transcriptional fusion of the omlA promoter with the sacB gene

Accession no. AJ868288

[19]

 pEMapxIIC

Conjugative vector pEMOC2 with a 270 bp deletion in the apxIIC gene which have a 1.4-kb upstream fragment and 1.4-kb downstream fragment

This work

Primers

 IICLF

5’ GCGTCGACATGACAACACCAATGATTGATTTAC 3’, upstream primer with internal SalI site (underlined)

This work

 IICLR

5’ AATCCCCGAA AGCATCATCCCTCCCATTC 3’, downstream primer with reverse complement sequence(underlined) of sequence in bold from primer IICRF

This work

 IICRF

5’ GGATGATGCT TTCGGGGATTCATCTCTATTG 3’, upstream primer with reverse complement sequence(underlined) of sequence in bold from primer IICLR

This work

 IICRR

5’ TTGCGGCCGCGTTGTAATAAGTCCCGTAACACCAG 3’, downstream primer with internal NotI site (underlined)

This work

 IICJDF

5’ GAAGAGCCATTACCCAACAAC 3’, upstream primer for identification of apxIIC gene deletion

This work

 IICJDR

5’ ATACAATAGAGATGAATCCCCG 3’, downstream primer for identification of apxIIC gene deletion

This work