Table 4.
Genetic engineering tools used in various Komagataeibacter strains
| Induction of gene expression | ||||
|---|---|---|---|---|
| Vector backbone | Promoters | Derived constructs | Komagataeibacterstrain | References |
| pSA19 | lac |
pSA-SD pSA-SD-S11E |
K. sucrofermentans BPR 2001 | (Nakai et al. 1999) |
| pBBR122 |
Bla lac |
pBla-Vhb-122 pLacDAAO-122 pBla-Vhb-122 pBBR-Glc-NAc |
K. xylinus BCRC 12334 K. xylinus BCRC 12334 K. xylinus CGMCC 2955 K. xylinus 10245 |
(Setyawati et al. 2007) (Setyawati et al. 2009) (Liu et al. 2018b) (Yadav et al. 2010) |
| pTI99 | trc |
pTI99-cdrS pTI99-motAB −motA; −motB |
K. hansenii ATCC 23769 K. hansenii ATCC 23769 K. hansenii ATCC 53582 |
(Fang et al. 2015) (Jacek et al. 2018) (Jacek et al. 2018) |
| pTSa |
tac gapA |
pTSaEX1 −pfk −pgi; −pgk; −fba; −tpi; −gap; −gpm; −pck; −mae; pTSaEX2 |
K xylinus DSM 2325 | (Gwon et al. 2019) |
| pIN01 | tac | pIN01-crp | K xylinus DSM 2325 | (Gwon et al. 2019) |
| pSEVA331Bb |
J23104 lux tet Bad J23100 J23101 J23102 J23104 J23105 J23108 J23109 J23110 J23115 J23119 J23119-A27T |
J-sRNA-331Bb pLux01 pLux02 pTet01 pTet02 pReceiver pSender pSEVA331Bb-mRFP pSEVA331Bb-NAG5-AGM1-UAP1 |
K. rhaeticus iGEM K. rhaeticus iGEM K. rhaeticus iGEM K. hansenii ATCC 53582 K. xylinus ATCC 700178 |
(Florea et al. 2016) a (Walker et al. 2018) (Teh et al. 2019) |
| pLBT | Promoter-less miniTn10 | pLBT::lacZ:kan | K. xylinus ITDI | (Battad-Bernardo et al. 2004) |
| Knock-out or inhibition of gene expression | ||||
| Vector backbone | Method/selection marker | Obtained constructs | Komagataeibacterstrain | References |
| pUC18 |
HR/ampR HR/kanR |
pUCA-EP | K. sucrofermentans BPR 2001 | (Ishida et al. 2002) |
| acsD deletion | K. hansenii ATCC 23769 | (Hu et al. 2010) | ||
| pUC19 | HR/ampR | T-GDH-Amp | K. xylinus BCRC 12334 | (Kuo et al. 2015) |
| pT7Blue | HR/cat-1 | pT7-Blue-PGDHC-I | K. sucrofermentans BPR 2001 | (Shigematsu et al. 2005) |
| pET14B | HR/kanR | motA motB and motAB disruption | K. hansenii ATCC 53582 | (Jacek et al. 2019b) |
| pHSG399 |
HR kanR HR ampR |
Dgc1 cmcAx pSA-ORF2/k (cpcAx deletion) |
K. sucrofermentans BPR 2001 |
(Bae et al. 2004) (Nakai et al. 2013) (Nakai et al. 2002) |
| pSEVA331Bb | sRNA | J-sRNA-331Bb (co-expression of Hfq and sRNA targeting UGPase mRNA) | K. rhaeticus iGEM | (Florea et al. 2016) |
| ter |
ECK120033736 ECK120010818 ECK120010799 ECK120051401 BBa_B0010 L3S2P21 L3S3P21 L3S2P24 L3S2P44 L3S1P47 |
K. rhaeticus iGEM K. hansenii ATCC 53582 K. xylinus ATCC 700178 |
(Teh et al. 2019) | |
| Protein degradation tags |
LAA AAV LVA DAS DAS + 2 |
K. rhaeticus iGEM K. hansenii ATCC 53582 K. xylinus ATCC 700178 |
||
| CRISPRi | pSEVA331Bb-Cas9-3xFLAG-sgRNA | K. hansenii ATCC 53582 | ||
HR homologous recombination, CRISPRi deactivated Caspase9-based system = CRISPR interference, sRNA short RNA mediated, ter translation terminator, kanR kanamycin resistance gene, cat-1 chloramphenicol acetyltransferase gene, ampR ampicillin resistance gene
aOther vector backbones reported here as replicating in K. rhaeticus iGEM but not studied further: pBla-Vhb-122; pSEVA321Bb, pBAV1K-T5-sfGFP