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Table 1 Strains, plasmids, and primers used in this study

From: Identification and characterization of a novel stress-responsive outer membrane protein Lip40 from Actinobacillus pleuropneumoniae

Strain, plasmid and primer Relevant characteristics Source
A. pleuropneumoniae
SLW01 Serovar 1 Weicheng Bei
13039 Serovar 10 Pat Blackall
E. coli
DH5a Cloning vehicle: supE44 ΔlacU169 (φ80 lacZΔM15) hsdR17recA1 endA1 gyrA96 thi-1 relA1 Takara (Dalian, China)
BL21(DE3) Expression host: F ompT r B m B; DE3 is a λ derivative carrying lacI and T7 RNA polymerase genes under placUV5 control Takara (Dalian, China)
Plasmid
pMD18-T E. coli cloning vector carrying an ampicillin resistance determinant Takara (Dalian, China)
pMD-lip40 pMD18-T carrying lip40 gene of A. pleuropneumoniae strain SLW01. This work
pGEX-KG N-terminal glutathione S-transferase (GST) fusion expression vector: pBR322 ori, Ampr [19]
pGEX-lip40 pGEX-KG carrying the lip40 gene for over-expression Lip40 protein This work
Primers (Primers were all synthesized in Sangon, Shanghai, China)
P1 5′ ATG AAA AAC ATC ACA AAA TTT GCA G 3′, upstream primer for lip40 gene cloning This work
P2 5′ TTA CTT TTG TTG TTT TGC GCC AAA 3′, downstream primer for lip40 gene cloning This work
P3 5′ CGG TTC GAT TTG GTG TGT ATG A 3′,upstream primer of lip40 gene for qRT-PCR analysis This work
P4 5′ AAC AAG TAA GCA TCA CCT GTG T 3′, downstream primer of lip40 gene for qRT-PCR analysis This work
P5 5′ AAG TGG CAG AGC TGG AAG AT 3′, upstream primer of internal control gene rluC for qRT-PCR analysis This work
P6 5′ TCA CAC CAA AAC TCA AGC CG 3′, downstream primer of internal control gene rluC for qRT-PCR analysis This work
P7 5′ TTG GAT CCT GTG GCA GTA AGA ACC ATT C 3′, upstream primer with BamHI site (underlined) comprising positions 58 to 77 of lip40 coding sequence This work
P8 5′ GGA AGC TTT TAC TTT TGT TGT TTT GCG C 3′, downstream primer with HindIII site (underlined) comprising positions 878 to 897 of lip40 coding sequence This work