Table 2 Primers used in this study
Primer Name | Primer Sequences (5′ to 3′) | Purposes |
|---|---|---|
BamHI-SaEsxA-F | CGGGATCCATGGCAATGATTAAGATG | amplify SaEsxA-his |
NotI-SaEsxA-his-taa-R | ATAAGAATGCGGCCGCTTAATGATGATGATGATGATGTTGCAAACCGAAATTATTAGAAAGTTGTTG | amplify SaEsxA-his |
HindIII-SaEsXB-F | CCCAAGCTTGGTGGATATAAAGGTATTAAAGCAGATG | amplify SaEsxB-his |
NotI-SaEsxB-his-taa-R | ATAAGAATGCGGCCGCTTAATGATGATGATGATGATGTGGGTTCACCCTATCAAGCC | amplify SaEsxB-his |
XhoI-PagC-F | CCGCTCGAGGTTAACCACTCTTAATAATAATGGGTTTTATAGC | amplify PpagC |
PacI-PagC-R | CCTTAATTAATACTACTTATTATTTACGGTGTGTTTAAACAC | amplify PpagC |
check-pCASP-R | CCGCCTTTGAGTGAGCTGATAC | check sequence from the down stream of heterologous genes |
check-PsicA-F | CGATCAACGTCTCATTTTCGCC | check sequence from the upstream of PsicA |
XhoI-PsicA-F | CTCGAGCCACAAGAAAACGAGG | amplify PsicA |
PsicA-R (no sicA rbs) | CACCGACTTTGTAGAACTTAACG | amplify PsicA |
(PsicA)invB-F | CGGTGACAGATAACAGGAGTAAGTATTAATTAAGGAAAAGATCTATGCAACATTTGG | amplify (PsicA)-invB |
invB-R (sipA) | CACTTGTAACCATTATTAATATCCTCTTCTGTTATCTCATTAGCGACCGACTAAAAAC | amplify (PsicA)-invB-(sipA) |
invB-R(sopE2) | CATTTTCTCCTCTTTAATTTATCTCATTAGCGACCGACTAAAAAC | amplify (PsicA)-invB-(sopE2) |
invB-R | ATCTCATTAGCGACCGACTAAAAAC | amplify (PsicA)-invB |
(invB)-sipA-F | GTTTTTAGTCGGTCGCTAATGAGATAACAGAAGAGGATATTAATAATGGTTACAAGTG | amplify (invB)-sipA |
HindIII-sipA-R | CCCAAGCTTTCCTGACTGAAAATACAAATTCTCTCCACCGCCAGTGTTATTTTTGATAATATCTAAC | amplify (invB)-sipA |
(invB)-sopE2-F | GATAAATTAAAGAGGAGAAAATGACTAACATAACACTATCCACCCAG | amplify (invB)-sopE2 |
HindIII-sopE2-R | AAGCTTTCCTGACTGAAAATACAAATTCTCTCCGGCCGGATCTTTACTCGC | amplify (invB)-sopE2 |
(invB)-sopA-F | GTTTTTAGTCGGTCGCTAATGAGATaaTTGATAAGGAATTGTAATGAAGATATCATCAGG | amplify (invB)-sopA |
HindIII-sopA-R | AAGCTTTCCTGACTGAAAATACAAATTCTCTCCCTTGCCTGCATTATTTGTATCTTTAATATTTTTAAC | amplify (invB)-sopA |
(sicA)-sipC-F | GTGAACAAGAAAAGGAATAATAAAGGGAGAAAAATATGTTAATTAGT | amplify (sicA)-sipC |
HindIII-sipC-R | AAGCTTTCCTGACTGAAAATACAAATTCTCTCCTCCGCTAATATCAAAAAACTTTCCGAC | amplify (sicA)-sipC |
(PsicA)sigE-F | CGTTAAGTTCTACAAAGTCGGTG GAGTCTTGAGGTAACTATATGGAAAGTC | amplify (PsicA)-sigE-(sopB) |
sigE-R-(sopB) | CCTGATTATGCATAATGCTCTTTCAATTGCTTC | amplify (PsicA)-sigE-(sopB) |
(sigE)sopB-F | GAGCATTATGCATAATCAGGAATATTAAAAACGCTATGCAAATAC | amplify (sigE)-sopB |
HindIII-sopB-R | AAGCTTTCCTGACTGAAAATACAAATTCTCTCCGTTATTAAGCTGCTTGACCTGAGC | amplify (sigE)-sopB |
pyrF-5’ | ATCCAATTTGCGCCACTTCCGGTGCCCATCATCAAGAAGGTCTGGTCATGCCGATCATATTCAATAACCCT | knockout pyrF gene |
pyrF-3’ | CCCCGTCTGCGTTGAATAAACCAGACGACTATTGGAATCGCTCATTATGCGACTAGTGAACCTCTTCGAGGG | knockout pyrF gene |
check-pyrF-R | CGGTATCGTTGTCAGAAATGCGGT | check pyrF deletion |
check-pyrF-R | CGTGATTGGTCACCAGGTTGGAAA | check pyrF deletion |
invA-5’ | GCAGAACAGCGTCGTACTATTGAAAAGCTGTCTTAATTTAATATTAACAGGATACCTATAGCCGATCATATTCAATAACCCT | knockout invA gene |
invA-3’ | CGGAACGAACTAATTCAGCGATATCCAAATGTTGCATAGATCTTTTCCTTAATTAAGCCCGACTAGTGAACCTCTTCGAGGG | knockout invA gene |
check-invA-F | TTACCAAAGCGTTTAATGCG | check invA deletion |
check-invA-R | CATCCTTCCATTATGGTCAT | check invA deletion |