Table 2.
Primers used to amplify gene fragments
| Gene | Template | Primer name | Sequencea | Restriction enzyme site |
|---|---|---|---|---|
| LysM | Genomic DNA of B. subtilis 168 (ATCC 23857) | LysM_For | GAGCAGCTCGAGGCACAAAGCATTAAGGTGAAAAAAGG | XhoI |
| LysM_Rev_Flag | GCTGCTGCTAGCTTATTATTTGTCATCGTCATCTTTATAATCGACTAACGCTTTTGCATCAGAAACCAGCTTG | NheI | ||
| LysM-mini-CipA | Genomic DNA of B. subtilis 168 (ATCC 23857) | LysM_For | GAGCAGCTCGAGGCACAAAGCATTAAGGTGAAAAAAGG | XhoI |
| LysM_Rev | GCTGCTGATATCGACTAACGCTTTTGCATCAGAAACC | EcoRV | ||
| Genomic DNA of C. thermocellum (ATCC 27405) | MC_For | GTAAGTAGATATCGTATCGGCGGCCACAATGACAGTCG | EcoRV | |
| MC_Rev_Flag | GCAGTAGCTAGCTTATTATTTGTCATCGTCATCTTTATAATCATTCGAATCATCTGTCGGTGTTGTTACAGG | NheI | ||
| pNWP43N | pNWP43N-BsCel5 | pNWP43N_For | GCCGACGCTAGCTTAAGCTTTTTTTTGGCGGACATCAGTAAC | NheI |
| PNWP43N_Rev | GACTATCTCGAGACCTGCAGCTGAGGCATGTGTTACAAAAAC | XhoI | ||
| Mini-CipA | Genome of Clostridium thermocellum | Mini-CipA_For | GTAGTACATATGGTATCGGCGGCCACAATGACAG | NdeI |
| Mini-CipA_Rev | GCAGTACTCGAGATTCGAATCATCTGTCGGTGTTG | XhoI | ||
| BsCel5′ | Genome of Bacillus subtilis | BsCel5_For | CCTCAGCATATGGCAGGGACAAAAACGCC | NdeI |
| BsCel5′_Rev | CTCCGGTTCTTCTGGGTCTACTCCTCCAGAAATACCATTT TCCTGTGTGGGTTTATC | |||
| Genome of Clostridium thermocellum | DocK_For | GGAGGAGTAGACCCAGAAGAACCGGAGGTTATTTATG | ||
| DocK_Rev | GCCGCCCTCGAGTTTATGTGGCAATACATCTATC | XhoI | ||
| CtCel9 | Genome of Clostridium thermocellum | CtCelF_For | GCTTCACATATGGCGGATTTCAACTATGGTGAGGCAC | NdeI |
| CtCelF_Rev | GGACCATCTCGAGCTGTTCAGCCGGGAATTTTTCAATAAG | XhoI | ||
| CpCel48 | pP43N-Cpcel48 | Cpcel48_For | CCTCTGCATATGGGTGAAACTGAGCAAGC | NdeI |
| Cpcel48_Rev | GTAGAGGACCCACCTCCTCCAGATCCTGGTTCGATACCCCAATTAAGTTTTCC | |||
| Genome of Clostridium thermocellum | DocS_For | GGATCTGGAGGAGGTGGGTCCTCTACTAAATTATACGGCGACGTC | ||
| DocS_Rev | GCATTACTCGAGGTTCTTGTACGGCAATGTATC | XhoI |
a
Restriction enzyme sites included in primer sequences for cloning purposes are indicated in bold, the Flag tag sequences are indicated by underlining, and the overlapping sequences are indicated by italics.