Table 1. L. monocytogenes strains, plasmids and primers used in the present study.
Strains 10403S, DP-L1935 and DP-L2787 have been described previously [10,14,15]. All of the other strains and plasmids were generated for the present study. List of plasmids refer to pPL2 constructs that were made to generate the chromosomal integrants expressing wild-type PC-PLC and PC-PLC propeptide mutants. Primer numbers refer to primers used to generate each construct. For primer sequences, see Supplementary Table S1 at http://www.BiochemJ.org/bj/432/bj4320557add.htm.
| Strain | Genotype and relevant features | Plasmid | Primer |
|---|---|---|---|
| 10403S | Wild-type strain | ||
| DP-L1935 | Internal in-frame deletion of plcB in 10403S background | ||
| DP-L2787 | Internal in-frame deletions of mpl and plcB in 10403S background | ||
| HEL-1031 | DP-L1935 pPL2:PactA–plcB | pERS1018 | 512–515 |
| HEL-1034 | DP-L1935 pPL2:PactA–plcB C28A C29A D30A | pERS1025 | 516, 517 |
| HEL-1037 | DP-L1935 pPL2:PactA–plcB E31A Y32A L33A | PERS1026 | 518, 519 |
| HEL-1040 | DP-L1935 pPL2:PactA–plcB P39A H40A D41A | pERS1027 | 526, 527 |
| HEL-1043 | DP-L1935 pPL2:PactA–plcB ΔC28–P39 | pERS1028 | 522, 523 |
| HEL-1046 | DP-L1935 pPL2:PactA–plcB ΔH40–P47 | pERS1029 | 524, 525 |
| HEL-1056 | DP-L1935 pPL2:PactA–plcB Q34A T35A P36A | pAB1049 | 520, 521 |
| HEL-1058 | DP-L1935 pPL2:PactA–plcB A37Q A38Q | pAB1050 | 528, 529 |
| HEL-1060 | DP-L1935 pPL2:PactA–plcB I42A D43A S44A | pAB1051 | 530, 531 |
| HEL-1062 | DP-L1935 pPL2:PactA–plcB K45A L46A P47A | pAB1052 | 532, 533 |
| HEL-1064 | DP-L1935 pPL2:PactA–plcB H48A K49A | pAB1053 | 534, 535 |
| HEL-1066 | DP-L1935 pPL2:PactA–plcB L50A S51A | pAB1054 | 536, 537 |
| HEL-1068 | DP-L1935 pPL2:PactA–plcB Q34K T35P | pERS1055 | 538, 539 |
| HEL-1070 | DP-L2787 pPL2:PactA–plcB | pERS1018 | 513, 514 |
| HEL-1074 | DP-L2787 pPL2:PactA–plcB ΔC28–P39 | pERS1028 | 522, 523 |
| HEL-1106 | DP-L1935 pPL2:PactA–plcB D30A E31A | pAPB1101 | 555, 556 |
| HEL-1112 | DP-L1935 pPL2:PactA–plcB ΔC28–L33 | pAB1087 | 549, 550 |
| HEL-1114 | DP-L1935 pPL2:PactA–plcB ΔE31–P36 | pAB1089 | 553, 554 |
| HEL-1116 | DP-L1935 pPL2:PactA–plcB ΔQ34–P39 | pAB1088 | 551, 552 |
| HEL-1134 | DP-L1935 pPL2:PactA–plcB ΔC28–T35 | pERS1123 | 570, 571 |
| HEL-1136 | DP-L1935 pPL2:PactA–plcB ΔC28–A37 | pERS1124 | 572, 573 |
| HEL-1138 | DP-L1935 pPL2:PactA–plcB ΔA38–K49 | pERS1125 | 574, 575 |
| HEL-1140 | DP-L1935 pPL2:PactA–plcB ΔC28–D41 | pERS1126 | 576, 577 |
| HEL-1142 | DP-L1935 pPL2:PactA–plcB ΔC28–D43 | pERS1127 | 578, 579 |
| HEL-1144 | DP-L1935 pPL2:PactA–plcB ΔC28–K45 | pERS1128 | 580, 581 |
| HEL-1146 | DP-L1935 pPL2:PactA–plcB ΔC28–S51 | pERS1129 | 582, 583 |
| HEL-1161 | DP-L1935 pPL2:PactA–plcB Q34A T35A | pERS1157 | 597, 598 |
| HEL-1164 | DP-L1935 pPL2:PactA–plcB P36A | pERS1158 | 599, 600 |
| HEL-1166 | DP-L1935 pPL2:PactA–plcB Y32A | pERS1159 | 601, 602 |
| HEL-1167 | DP-L1935 pPL2:PactA–plcB L33A | pERS1160 | 603, 604 |
| HEL-1169 | DP-L2787 pPL2:PactA–plcB ΔC28–L33 | pAB1087 | 549, 550 |
| HEL-1176 | DP-L2787 pPL2:PactA–plcB ΔC28–T35 | pERS1123 | 570, 571 |
| HEL-1177 | DP-L2787 pPL2:PactA–plcB ΔC28–A37 | pERS1124 | 572, 573 |
| HEL-1182 | DP-L2787 pPL2:PactA–plcB ΔC28–D41 | pERS1126 | 576, 577 |
| HEL-1184 | DP-L2787 pPL2:PactA–plcB ΔC28–D43 | pERS1127 | 578, 579 |
| HEL-1185 | DP-L2787 pPL2:PactA–plcB ΔC28–K45 | pERS1128 | 580, 581 |
| HEL-1187 | DP-L2787 pPL2:PactA–plcB ΔC28–S51 | pERS1129 | 582, 583 |
| HEL-1204 | DP-L2787 pPL2:PactA–plcB ΔC28–H48 | pERS1189 | 609, 610 |
| HEL-1206 | DP-L2787 pPL2:PactA–plcB ΔC28–L50 | pERS1190 | 611, 612 |
| HEL-1223 | DP-L1935 pPL2:PactA–plcB E31A | pAB1217 | 595, 596 |
| HEL-1320 | DP-L1935 pPL2:PactA–plcB ΔC28–H48 | pERS1189 | 609, 610 |
| HEL-1321 | DP-L1935 pPL2:PactA–plcB ΔC28–L50 | pERS1190 | 611, 612 |