Table 1.
Plasmids and strains used in this study.
| Plasmid or Straina | Relevant genotypeb |
|---|---|
| ssrA-tagging vectors | |
| pKG1268 | pGEMcat-(Ec)ssrA* |
| pKG1519 | pGEMcat-(Cc)ssrA-ALGG |
| pKG1520 | pGEMkan-(Ec)ssrA* |
| pKG1521 | pGEMkan-(Cc)ssrA-ALGG |
| pKG1522 | pGEMkan-(Cc)ssrA(A2D)-ALGG |
| pKG1523 | pGEMcat-(Cc)ssrA(A2D)-ALGG |
| pKG1524 | pGEMcat-HA-(Ec)ssrA* |
| pKG1525 | pGEMcat-his6-(Ec)ssrA* |
| pKG1526 | pGEMcat- myc -(Ec)ssrA* |
| pKG1527 | pGEMcat-HA-(Cc)ssrA(A2D)-ALGG |
| pKG1528 | pGEMcat-his6-(Cc)ssrA(A2D)-ALGG |
| pKG1529 | pGEMcat-myc-(Cc)ssrA(A2D)-ALGG |
| pKG1532 | pGEMcat-GFP-(Ec)ssrA* |
| SspB expression strains | |
| KG844 | amyE∷{Pspank-(Ec)sspB spc} (pKG1266) |
| KG1030 | amyE∷{Pspank-(Cc)sspB spc} |
| KG1096 | lacA∷{Pxyl-(Ec)sspB tet} (pKG1267) |
| KG1098 | amyE∷{Pspank(T-7A)-(Ec)sspB spc} (pKG1334) |
| KG1100 | amyE∷{Pspank(T-35C)-(Ec)sspB spc} (pKG1457) |
| Characterization of modified ssrA tags | |
| KG854 | thrC∷{Pc-gfp-(Ec)ssrA-ALDD+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG858 | thrC∷{Pc-gfp-(Ec)ssrA-ALAA+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG863 | thrC∷{Pc-gfp erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG884 | thrC∷{Pc-gfp-(Ec)ssrA-ALWW+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG885 | thrC∷{Pc-gfp-(Ec)ssrA-AISS+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG886 | thrC∷{Pc-gfp-(Ec)ssrA-AISV+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG887 | thrC∷{Pc-gfp-(Ec)ssrA-ALQS+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG888 | thrC∷{Pc-gfp-(Ec)ssrA-ADAS+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG889 | thrC∷{Pc-gfp-(Ec)ssrA-ADAT+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG890 | thrC∷{Pc-gfp-(Ec)ssrA-ADAC+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG892 | thrC∷{Pc-gfp-(Ec)ssrA-ADCS+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG893 | thrC∷{Pc-gfp-(Ec)ssrA-ADAN+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG919 | thrC∷{Pc-gfp-(Ec)ssrA-ADVS+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG920 | thrC∷{Pc-gfp-(Ec)ssrA-ADTS+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG922 | thrC∷{Pc-gfp-(Ec)ssrA-ADSA+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG923 | thrC∷{Pc-gfp-(Ec)ssrA-ADAG+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG924 | thrC∷{Pc-gfp-(Ec)ssrA-ADAV+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG925 | thrC∷{Pc-gfp-(Ec)ssrA-ADKS+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG926 | thrC∷{Pc-gfp-(Ec)ssrA-AQSA+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG927 | thrC∷{Pc-gfp-(Ec)ssrA-AHHA+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG946 | thrC∷{Pc-gfp-(Ec)ssrA* erm clpC∷tet} amyE∷{Pspank-(Ec)sspB spc} |
| KG951 | thrC∷{Pc-gfp-(Ec)ssrA* erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG952 | thrC∷{Pc-gfp-(Ec)ssrA-ADSS+4 erm} amyE∷{Pspank-(Ec)sspB spc} |
| KG1060 | comA∷pGEMcat-comA-(Ec)ssrA* cat (pKG1458) amyE∷{Pspank-(Ec)sspB spc} |
| KG1068 | thrC∷{Pc-gfp-(Bs)ssrA-ALGG erm} |
| KG1185 | thrC∷{Pc-gfp-(Ec)ssrA-AHHA+4 erm clpX∷spec∷kan} |
| srfA-lacZ strains | |
| KG1178 | amyE∷{Pspank-(Ec)sspB spc} comA∷pGEMcat-comA-(Ec)ssrA* cat (pKG1458) thrC∷{PsrfA-lacZ erm} |
| KG1177 | amyE∷{Pspank(T-7A)-(Ec)sspB spc} comA∷pGEMcat-comA-(Ec)ssrA* cat (pKG1458) thrC∷{PsrfA-lacZ erm} |
| KG1180 | amyE∷{Pspank(T-35C)-(Ec)sspB spc} comA∷pGEMcat-comA-(Ec)ssrA* cat (pKG1458) thrC∷{PsrfA-lacZ erm} |
| KG1205 | comA∷pGEMcat-comA-(Ec)ssrA* cat (pKG1458) thrC∷{PsrfA-lacZ erm} |
| KG1233 | comA∷pGEMcat-comA-(Ec)ssrA-ADAT+4 cat (pKG1461) thrC∷{PsrfA-lacZ erm} |
| KG1234 | comA∷pGEMcat-comA-(Ec)ssrA-DDAS+4 cat thrC∷{PsrfA-lacZ erm} |
| KG1235 | comA∷pGEMcat-comA-(Ec)ssrA-ADAN+4 cat (pKG1460) thrC∷{PsrfA-lacZ erm} |
| KG1343 | comA∷pGEMcat-comA (wild type) cat (pKG1459) thrC∷{PsrfA-lacZ erm} |
| Sporulation mutants | |
| AG522 | kinA∷Tn917 |
| NY120 | ΔkinB∷spc |
| AG1468 | Δspo0J∷spc |
| KG1061 | amyE∷{Pspank-(Ec)sspB spc} spo0J ∷ pGEMcat-spo0J-(Ec)ssrA* cat (pKG1465) |
| KG1071 | amyE∷{Pspank-(Ec)sspB spc} kinA∷pGEMcat-kinA-(Ec)ssrA* cat (pKG1463) |
| KG1073 | amyE∷{Pspank-(Ec)sspB spc} kinB∷pGEMcat-kinB-(Ec)ssrA* cat (pKG1464) |
| (Ec)SspB and (Cc)SspB discrimination | |
| KG1018 | amyE∷{Pspank-(Ec)sspB spc} thrC∷{Pc-gfp-(Cc)ssrA-ALGG erm} |
| KG1019 | amyE∷{Pspank-(Ec)sspB spc} thrC∷{Pc-gfp-(Cc)ssrA-ALDD erm} |
| KG1020 | amyE∷{Pspank-(Ec)sspB spc} thrC∷{Pc-gfp-(Cc)ssrA(A1W)-ALGG erm} |
| KG1021 | amyE∷{Pspank-(Ec)sspB spc} thrC∷{Pc-gfp-(Cc)ssrA(A2D)-ALGG erm} |
| KG1023 | amyE∷{Pspank-(Cc)sspB spc} thrC∷{Pc-gfp-(Ec)ssrA* erm} |
| KG1024 | amyE∷{Pspank-(Cc)sspB spc} thrC∷{Pc-gfp-(Ec)ssrA-ALDD+4 erm} |
| KG1025 | amyE∷{Pspank-(Cc)sspB spc} thrC∷{Pc-gfp-(Cc)ssrA-ALGG erm} |
| KG1026 | amyE∷{Pspank-(Cc)sspB spc} thrC∷{Pc-gfp-(Cc)ssrA-ALDD erm} |
| KG1027 | amyE∷{Pspank-(Cc)sspB spc} thrC∷{Pc-gfp-(Cc)ssrA(A1W)-ALGG erm} |
| KG1028 | amyE∷{Pspank-(Cc)sspB spc} thrC∷{Pc-gfp-(Cc)ssrA(A2D)-ALGG erm} |
| KG1201 | amyE∷{Pspank-(Cc)sspB spc} lacA∷{Pxyl-(Ec)sspB tet thrC∷{Pc-gfp-(Ec)ssrA* erm} comA∷pGEMcat-comA-(Cc)ssrA(A2D)-ALGG cat (pKG1466) |
a
All B. subtilis strains are derivatives of JH642 and contain mutations in trpC and pheA (not shown).
b
(Ec)ssrA-ALGG+4 is referred to as (Ec)ssrA*. The plasmid name is in parenthesis used to create the single cross-over integrations.