Table 2 Primers used for construction and verification of pGB-sfGFP-best, pMM7, and the P. thermoglucosidasius ∆spo0A strain
No | Sequence 5ʹ-3ʹ | Description |
|---|---|---|
Geo Vector backbone-For | AGTTTGUTGAAGATTAGATGCTATAATTGTT | Forward primer to amplify pGB backbone from pMTL61110 |
Geo Vector backbone-Rev | AGCCGGTTGUTTTGCCGGCCGGCCCTACTCTTTA | Reverse primer to amplify pGB backbone from pMTL61110 |
P7_sfGFP_Ter-For | ACAACCGGCUCCTTTTGCTCTTTCTTCT | Forward primer to amplify P7-sfGFP from pIP7 |
P7_sfGFP_Ter-Rev | ACAAACUGAGAAAAAGAAACAGAGGC | Reverse primer to amplify P7-sfGFP from pIP7 |
SeqIns_For | TTGCAATGTGGAATTGGG | Forward primer to amplify and sequence the sfGFP insert region in the vector |
SeqIns_Rev | ATAACTCGTCTTCCTAAGCATCC | Reverse primer to amplify and sequence the sfGFP insert region in the vector |
F11V/T13P-For | ATTCTGGUGGAACTGGATGGTGATGTC | Forward primer for introduction of F11V/T13P in sfGFP |
F11V/T13P-Rev | ACCAGAAUAGGGACGACACCAGTGAACAGCTCTTCGCC | Reverse primer for introduction of F11V/T13P in sfGFP |
N39D-For | ACTGATGGUAAACTGACGCTGAAGTTCATCTG | Forward primer for introduction of N39D in sfGFP |
N39D-Rev | ACCATCAGUTGCGTCACCTTCACCCTC | Reverse primer for introduction of N39D in sfGFP |
A179A-For | AGCTGGCCGAUCACTACCAGCAAAACACT | Forward primer for introduction of A179A in sfGFP |
A179A-Rev | ATCGGCCAGCUGCACGCTGCCATCCTCCA | Reverse primer for introduction of A179A in sfGFP |
23 | ACCCGGGGUTCCTCTAG | Forward primer to amplify pMM7 backbone fragment from pGB-sfGFP-best |
24d | AATTCGUAATCATGGTCATATGGATACAGCG | Reverse primer to amplify pMM7 backbone fragment from pGB-sfGFP-best |
11M2 | GGCCGCTGTATCCATATGACCATG | Forward primer for sequencing of homologous arms region in pMM7 |
12 | GTTGTAAAACGACGGCCAGTGC | Reverse primer for sequencing of homologous arms region in pMM7 |
PNJ1241 | ACGAATUTAATATCACTCGATGCTTAAAATCAATCTCATCAACC | Forward primer to amplify spo0A left homologous arm from gDNA |
PNJ1242 | AGCCGCUAATATCAACGGTTTAAATGTCATG | Reverse primer to amplify spo0A left homologous arm from gDNA |
PNJ1243 | AGCGGCUCTATGCAAAAAAAGCTAAATTTATGACGTTTTTCACG | Forward primer to amplify spo0A right homologous arm from gDNA |
PNJ1244 | ATCCCCGGGUCCGCATTGGATTATATATCCGAGATTCTGC | Reverse primer to amplify spo0A right homologous arm from gDNA |
PNJ1245 | CAGTTCCGTATCTGTATGTCTGTCCTTAAATCG | Forward primer for sequencing of spo0A genomic region |
PNJ1246 | GGAAAAAAATCGAAAACATGGATGATCTTTCC | Reverse primer for sequencing of spo0A genomic region |