The laboratory has implemented high throughput methodologies for preparation of vectors suitable for: heterologous expression in E. coli and baculovirus systems, cell transfection and lentiviruses. Moreover, we have developed a pipeline for efficient automatic, multistep purification of proteins and complexes expressed in E. coli. The lab is equipped with a Janus pippeting station and an AKTA Express Twin chromatography system.
In addition to preparations of vectors and protein purification for our own projects we are open to collaborations.
People involved:
Katarzyna Kowalska, Krystian Stoduś
Vectors we use:
1. Vectors for protein expression in E. coli
| vector | tag | resistance |
| pET28 SUMO Kan | N: HisTAG-SUMO- | Kanamycin |
| pET28 SUMO Amp | N: HisTAG-SUMO- | Ampicillin |
| pET28 PP | N: HisTAG-PP- | Kanamycin |
| pET28 Streptag | N: StrepTAG- | Ampicillin |
| pET28 SUMO Kan, pET28 PP | ——————— | Kanamycin |
| pET28 SUMO Amp | ——————— | Ampicillin |
| pET28 SUMO Kan, pET28 PP | C: -HisTAG | Kanamycin |
| pET28 SUMO Kan | C: -HisTAG | Ampicillin |
| pET28 HisTEV | N: HisTAG-TEV- | Kanamycin |
| pET28 GST-TEV | N: GST-TEV- | Kanamycin |
| pET28 TEV-His | C: -TEV-HisTAG | Kanamycin |
2. Mammalian expression vectors (Invotrogen’s pcDNA5/FRT/TO backbone with inserted tags)
| vector | tag | resistance |
| pcDNA5FRTTO N:FLAG | FLAG-TEV- | Ampicillin |
| pcDNA5FRTTO N:EGFP | EGFP-TEV- | Ampicillin |
| pcDNA5FRTTO N:mCherry | mCherry-TEV- | Ampicillin |
| pcDNA5FRTTO C:FLAG | -TEV-FLAG | Ampicillin |
| pcDNA5FRTTO C:EGFP | -TEV-EGFP | Ampicillin |
| pcDNA5FRTTO C:mCherry | -TEV-mCherry | Ampicillin |
| pcDNA5FRTTO C:protA | -TEV-protA | Ampicillin |
3. Other vectors
| vector | tag | resistance | |||||||||||||||||||||
| Lenitvirus | |||||||||||||||||||||||
| pLVX-Tight-Puro | -TEV-EGFP | Ampicillin | |||||||||||||||||||||
| HIV SSF RFP WPRE | -TEV-EGFP | Ampicillin | |||||||||||||||||||||
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