Table 1.
Plasmids and strains used in this study.
| Name | Description | Reference |
|---|---|---|
| Strain | ||
| E. coli TOP10 | Host cells for plasmid construction | Invitrogen |
| WT | Wildtype C. cellulolyticum H10 | ATCC |
| WT-P | WT with pRNAi control plasmids | This study |
| WT-P-pta | WT with pRNAi-pta plasmids | This study |
| WT-P-ack | WT with pRNAi-ack plasmids | This study |
| WT-P-afp | WT with pFd-AFP plasmids | (Xu et al., 2015) |
| WT-G | WT with a chromosomal Fd::empty cassette | This study |
| WT-G-afp | WT with a chromosomal Fd::afp cassette | This study |
| LM | ΔldhΔmdh | (Li et al., 2012) |
| LM-P | LM with pRNAi plasmids | This study |
| LM-P-pta | LM with pRNAi-pta plasmids | This study |
| LM-G | LM with a chromosomal RNAi control | This study |
| LM-G-pta | LM with a chromosomal Fd::pta asRNA cassette | This study |
| Plasmid | ||
| pRNAi | CMPr in E. coli; TMPr in H10; Fd::empty cassette | (Xu et al., 2015) |
| pRNAi-pta | pRNAi derivative with a Fd::pta asRNA cassette | This study |
| pRNAi-ack | pRNAi derivative with a Fd::ack asRNA cassette | This study |
| pLyc045 | pRNAi derivative with 3198up-mlsR-Fd::empty-3198down | This study |
| pLyc046 | pRNAi derivative with 3198up-mlsR-Fd::pta asRNA-3198down | This study |
| pFd-AFP | pRNAi derivative with a Fd::afp cassette | (Xu et al., 2015) |
| pLyc048 | pRNAi derivative with 3198up-mlsR-Fd::afp-3198down | This study |
| pCas9n-3198D | pRNAi derivative with a cas9 nickase and a gRNA targeting the 3198D site | (Xu et al., 2015) |
| pCas9n-3198D with donor | pCas9n-3198D derivative with left arm-3P4::pta asRNA-right arm | This study |