Table 3 Plasmids and strains used in this study

Lactiplantibacillus plantarum WCFS1

Host strain

[30]

Lactiplantibacillus pentosus KW1

Host strain

This study

Lactiplantibacillus pentosus KW2

Host strain

This study

Escherichia coli Stellar

Subcloning strain

Takara Bio

KW1-pEV

L. pentosus KW1 harboring pEV

This study

KW1-Cyt-H56

L. pentosus KW1 harboring pSIP_CytH56

This study

KW1-NTTM-H56

L. pentosus KW1 harboring pSIP_KW1_NTTM_H56

This study

KW1-Lipo-H56

L. pentosus KW1 harboring pSIP_KW1-2_Lipo_H56

This study

KW1-LysM-H56

L. pentosus KW1 harboring pSIP_KW1_LysM_H56

This study

KW1-LPxTG-H56

L. pentosus KW1 harboring pSIP_KW1-2_LPxTG_H56

This study

KW2-pEV

L. pentosus KW2 harboring pEV

This study

KW2-Cyt-H56

L. pentosus KW2 harboring pSIP_CytH56

This study

KW2-NTTM-H56

L. pentosus KW2 harboring pSIP_KW2_NTTM_H56

This study

KW2-Lipo-H56

L. pentosus KW2 harboring pSIP_KW1-2_Lipo_H56

This study

KW2-LysM-H56

L. pentosus KW2 harboring pSIP_KW2_LysM_H56

This study

KW2-LPxTG-H56

L. pentosus KW2 harboring pSIP_KW1-2_LPxTG_H56

This study

Plasmids

Description

Reference

pEV

Ery^r; 256_rep; pSIP401 derivative; control plasmid (“empty vector”)

[68]

pSIP_1261_H56-DC

Ery^r; 256_rep; pSIP401 derivative containing the inducible P_sppA promoter fused to a gene construct encoding the lipoprotein anchor sequence from the gene lp_1261 followed by the sequence encoding H56 and a dendritic cell binding (DC) peptide [69].

This study

pJET1.2_ESAT-6-Rv2660c

Amp^r; pJET1.2 derivative containing a gene fragment encoding the fused M. tuberculosis derived antigens ESAT-6 and Rv2660c.

GenScript

pLp_1261AgE6-DC

Ery^r; 256_rep; pSIP401 derivative containing the inducible P_sppA promoter translationally fused to the lipoprotein anchor sequence derived from lp_1261 followed by a sequence encoding AgE6 fused to a DC-peptide.

[18]

pSIP_KW1_NTTM_H56

Ery^r; 256_rep; pSIP401 derivative containing the inducible P_sppA promoter translationally fused to an N-terminal transmembrane anchor (NTTM) derived from NGP02_02010 followed by the sequence encoding H56 and the DC-peptide.

This study

pSIP_KW2_NTTM_H56

Ery^r; 256_rep; pSIP401 derivative containing the inducible P_sppA promoter translationally fused to an N-terminal transmembrane anchor (NTTM) derived from LPKW2_12625 followed by the sequence encoding H56 and the DC-peptide.

This study

pSIP_KW1-2_Lipo_H56

Ery^r; 256_rep; pSIP401 derivative containing the inducible P_sppA promoter translationally fused to the N-terminal signal peptide and Lipoprotein anchor derived from NGP02_02310 from KW1 (identical to LPKW2_10785 in KW2) followed by the sequence encoding H56 and the DC-peptide.

This study

pSIP_KW1_LysM_H56

Ery^r; 256_rep; pSIP401 derivative containing the inducible P_sppA promoter translationally fused to the N-terminal signal peptide and LysM anchor derived from the gene NGP02_07240 followed by the sequence encoding H56 and the DC-peptide.

This study

pSIP_KW2_LysM_H56

Ery^r; 256_rep; pSIP401 derivative containing the inducible P_sppA promoter translationally fused to the N-terminal signal peptide and LysM anchor derived from the gene LPKW2_06475 followed by the sequence encoding H56 and the DC-peptide.

This study

pSIP_3050_H56_cwa3001

Ery^r; 256_rep; pSIP_3050_H1_cwa3001 derivative containing the inducible P_sppA promoter translationally fused to the N-terminal signal peptide derived from the gene lp_3050 followed by the sequence encoding H56 and a C-terminal LPxTG anchor from lp_3001.

This study

pSIP_3050_H1_cwa3001

Ery^r; 256_rep; pLp_3050DC-AgE6cwa2 [18] derivative containing the inducible P_sppA promoter translationally fused to the N-terminal signal peptide derived from lp_3050 followed by the sequence encoding H1 and a C-terminal LPxTG anchor from lp_3001.

Unpublished

pSIP_KW1-2_LPxTG_H56

Ery^r; 256_rep; pSIP401 derivative containing the inducible P_sppA promoter translationally fused to the N-terminal signal peptide and LPxTG anchor derived from NGP02_06930 (identical to KW2_01306 in KW2) followed by the sequence encoding H56.

This study

pSIP_CytH56

Ery^r; 256_rep; pSIP401 derivative containing the inducible P_sppA promoter translationally fused to H56 and the DC-peptide.

This study

pSIP403_mCherry

Ery^r; 256_rep; pSIP403 derivative, encoding red fluorescent protein mCherry under control of the inducible P_sppA promoter.

[16]