Table 1. Relevant strains and primers used in this study.
| Strain | Description | Source | |
|---|---|---|---|
| TIGR4 | Wild-type serotype 4 encapsulated strain, GentR | Laboratory Strain; Ingeborg Aaberge | |
| TIGR4Δcps | Serotype 4 acapsular strain. Cps locus replaced with SpecR gene | Laboratory Strain; Andrew Camilli | |
| TIGR4ΔpepN | Serotype 4 strain with the pepN gene (SP_0797) replaced with a CmR gene | This work | |
| TIGR4pepNFLAG | Serotype 4 strain with a FLAG epitope tag immediately upstream of the stop codon in the pepN gene, SmR | This work | |
| TIGR4pepNRevertant | Serotype 4 strain expresses the wild-type pepN gene in the ΔpepN genetic background, SmR | This work | |
| Primer Name | Sequence (5' to 3') | ||
| For generating ΔpepN mutant | |||
| F0 ΔpepN Sequencing primer |
GAGTTTTTTGACGAAGGG | ||
| R0 ΔpepN Sequencing primer |
CCTGCCCATAGCTATTAA | ||
| F1 ΔpepN For DNA construct |
GCACATGTCGTTACAGAA | ||
| R1 ΔpepN For DNA construct |
CATCAAGCTTATCGATACCGGCTTGCATAGTTTTCTCC | ||
| F2 ΔpepN For DNA construct |
GAAGGTTTTTATATTACAGCTCCAGTCGAAGCAGTTGTTCTT | ||
| R2 ΔpepN For DNA construct |
ATCCAAGCCCAAAAATCG | ||
| For generating pepNFLAG mutant | |||
| F0 pepNFLAG For sequencing |
TGATCGGAGATGAAATCG | ||
| R0 pepNFLAG For sequencing |
CAGCTGATGGAATTTCAC | ||
| F1 pepNFLAG For DNA construct |
CCTTTTAGCGGATTTGGT | ||
| R1 pepNFLAG For DNA construct |
TTTATCATCATCATCTTTATAATCTGCATTTCCGTATTGAAG | ||
| F2 pepNFLAG For DNA construct |
GATTATAAAGATGATGATGATAAATAAATAAGCCTAAAATAAAAAGAA | ||
| R2 pepNFLAG For DNA construct |
GTCCCCAGAAGTTAATCT | ||
GentR, gentamycin resistant; SpecR, spectinomycin resistant; CmR, chloramphenicol resistant; SmR, streptomycin resistant
Bolded text indicates sequences that are homologous to the CmR cassette
Underlined text indicate the FLAG epitope sequence