Bacteria
Escherichia coli
Strain/Type
E. coli XL1-Blue, K12 derivative
Genotype: recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F´ proAB lacIqZΔM15 Tn10 (Tetr)]
Characteristics e.g. sensitizing or toxic effect, resistance to antibiotics
XL-1 cells are tetracycline resistant. XL1-Blue cells are endonuclease (endA) deficient, which greatly improves the quality of miniprep
DNA, and are recombination (recA) deficient, improving insert stability. The hsdR mutation prevents the cleavage of cloned DNA by the
EcoK endonuclease system. The lacIqZΔM15 gene on the F´ episome allows blue-white color screening.recA1, endA1, gyrA96, thi-1, hsdR17, supE44, relA1, lac [F´ proAB lacIqZΔM15 Tn10 (TetR)]
Approved as biological safety measure if taken as recipient organism for genetic engineering?
yes
Genetically modified (GenTSV)
Risk group (BioStoffV)
1
Risk assessment
According to ZKBS (https://zag.bvl.bund.de/ecoli/detail.jsf?dswid=2647&dsrid=605&id=54)
Storage location of aliquots in the Biolab (just click Bearbeiten in the right corner of the header to add or change information in the table and use the menue in the left header to e.g. add a row)
| source | bacterial strain | freezing date | amount of bacteria per vial | host cells, stock was produced on | no. of aliquots | belongs to (full name) | rack/box in N2 tank or -80°° freezer and location (room, address) | comments |
|---|---|---|---|---|---|---|---|---|
| Agilent | XL-1B | 21.1.2016 | nd | 21.1.2016 | 1 | Stefanie Wedepohl | -80°C "Omega", 115.2 Shelf B Rack 6 Box 1 H1 | glycerol stock |
| Agilent | XL-1B | var. | nd | 21.1.2016 | var. | Stefanie Wedepohl | -80°C "Omega", 115.2 ? |
Background
Used for production of recombinant proteins under the control of T7 RNA polymerase.
The strain contains DE3, a λ prophage carrying the T7 RNA polymerase gene under control of the lac UV5 promoter and lacIq. IPTG is required to induce expression of the T7 RNA polymerase. Protein expression from transforming plasmids containing a T7 promoter-driven expression system is repressed until IPTG induction of T7 RNA polymerase expression occurs. The strain does not contain the lon protease and is also deficient in the outer membrane protease, OmpT. The lack of two key proteases reduces the degradation of heterologous proteinsexpressed in the strain.
Shelf B Rack 6 Box 1 row 3 | varying number of aliquots of chemically competent cells prepared from glycerol stock | |||||||
| Stefanie Wedepohl | XL-1B | 100 µL | 25 | Robyn Diehn | -80°C "Omega", 115.2, B-2-2 | chemical competent | ||
| Stefanie Wedepohl | XL-1B | 4 | Robyn Diehn | -80°C "Omega", 115.2, B-2-5 | stock | |||
| Stefanie Wedepohl | XL-1B | 1 mL | 4 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV4_NT5B | ||
| Stefanie Wedepohl | XL-1B | 1 mL | 4 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV4_O-glyko_620_direkt | ||
| Stefanie Wedepohl | XL-1B | 1 mL | 4 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV4_O-glyko_1800_direkt | ||
| Stefanie Wedepohl | XL-1B | 1 mL | 05.12.23 | 3 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV5.1_D3_CO | |
| Stefanie Wedepohl | XL-1B | 1 mL | 05.12.23 | 3 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV5.4_NT5B_delta cys 1088/1130 ala Kol 5 | |
| Stefanie Wedepohl | XL-1B | 1 mL | 12.12.23 | 3 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV5.4_D3_delta cys 1088/1130 ala | |
| Stefanie Wedepohl | XL-1B | 1 mL | 12.12.23 | 3 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV5.4_o-glyko 620_6x His | |
| Stefanie Wedepohl | XL-1B | 1 mL | 26.01.24 | 4 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV4_O-glyko_620_direkt | |
| Stefanie Wedepohl | XL-1B | 1 mL | 26.01.24 | 4 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV4_O-glyko_1800_direkt | |
| Stefanie Wedepohl | XL-1B | 1 mL | 26.01.24 | 4 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV4_Mini 2 | |
| Stefanie Wedepohl | XL-1B | 200 µL | 21.03.24 | 8 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV4_D3_CO | |
| Stefanie Wedepohl | XL-1B | 200 µL | 21.03.24 | 8 | Robyn Diehn | -80°C "Omega", 115.2, B-2-17 | With Plasmid: SekTagV4_D3_CO delta cys 1088/1130 ala | |
| Stefanie Wedepohl | XL-1B | 1 mL | 4 | Robyn Diehn | -80°C "Omega", 115.2, B-2-8 | Stock | ||
| Stefanie Wedepohl | XL-1B | 1 mL | 05.03.24 | 3 | Robyn Diehn | -80°C "Omega", 115.2, B-2-8 | Stock with Tetracycline grown | |
Background
nalidixic acid resistant
tetracycline resistant (carried on the F plasmid)
Cultivation and freezing protocols
| View file | ||||
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Data sheets, further information
View file name XL-1B.pdf height 250
