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Table 1 Bacterial strains and plasmids used in this study

From: Production, secretion and purification of a correctly folded staphylococcal antigen in Lactococcus lactis

(A) Bacterial strains
Names Genotype, characteristics Reference
Escherichia coli
JM109 endA1, recA1, gyrA96, thi, hsdR17 (r k , m +k ), relA1, supE44, Δ(lac-proAB), [F´ traD36, proAB, laqIqZΔM15] Promega
Lactococcus lactis
MG1363 plasmid free derivative of NCDO712 Laboratory collection
S. aureus
Lowenstein (ATCC 49521) clinical isolate, capsular polysaccharide CP5-positive Laboratory collection
COL clinical isolate, methicillin resistant Laboratory collection
htrA 1 COL htrA 1 :: cat, CmR [57]
htrA 2 COL htrA 2 :: spc, SpcR [57]
(B) Plasmids
Names Characteristics Reference
pGEMT T-tailed PCR product Cloning vector, AmpR Promega
pVE8124 pGEMT derivative where recombinant htrA 1r ORF is cloned
htrA 1r encodes HtrA1-ΔTM-Ser255Ala-His6 protein
This work
pVE8125 pGEMT derivative where recombinant htrA 2r ORF is cloned
htrA 2r encodes SPExp4-HtrA2-ΔTM-Ser619Ala-His6 protein
This work
pLB145 pWV01; CmR; expression-secretion vector where exp4 SP -nuc is cloned under the control of a lactococcal expression system, P zit zitR
exp4 SP -nuc encodes a hybrid protein between a lactococcal signal peptide (SPExp4) and the mature secreted form of the staphylococcal nuclease (NucB)
[4]
pVE8126 pLB145 derivative where htrA 1r ORF is cloned in place of nuc ORF to be fused in frame to exp4 SP
exp4 SP -htrA 1r encodes a hybrid precursor leading to secreted rHtrA1 protein
This work
pVE8127 pLB145 derivative where htrA 2r ORF is cloned in place of exp4 SP -nuc ORF
htrA 2r encodes a hybrid precursor leading to secreted rHtrA2 protein
This work
  1. Amp R ampicillin resistant, Cm R chloramphenicol resistant, Spc R spectinomycin resistant.