LEXSY host T7-TR

For general laboratory use. Not intended for human or animal diagnostic or therapeutic uses.

Shipping: shipped on dry ice

Storage Conditions: store at -80 °C
avoid freeze/thaw cycles
Upon arrival the glycerol stocks must be stored at -80 °C or inoculated into LEXSY BHI.

Shelf Life: 12 months

Description:
The Leishmania tarentolae strain T7-TR is used as host strain for inducible LEXSY expression vectors.

Content:
3 vials with 1.6 ml each of frozen glycerol stocks of LEXSY host T7-TR (recombinant Leishmania tarentolae strain expressing bacteriophage T7 RNA polymerase and TET repressor).

These stocks can be stored at -80 °C for at least 1 year. For reactivation see below.

Organism:
Recombinant Leishmania tarentolae strain constitutively expressing bacteriophage T7 RNA polymerase and TET repressor.

Biosafety level:
1, Non-pathogenic for mammalians

Source:
Tarentola annularis

Reactivation of LEXSY host:
Thaw glycerol stock on ice and inoculate the entire content of the vial into 10 ml of LEXSY BHI medium (Cat.-No. ML-411) containing the antibiotics Nourseothricin (Cat.-No. AB-101) and LEXSY Hygro (Cat.-No. AB-104). Incubate at 26 °C and dilute as required.

Preparation of LEXSY BHI growth medium:
Dissolve 37 g/l LEXSY BHI powder (Cat.-No. ML-412) in deionized water and autoclave for 15 min at 121 °C. Add Hemin and PenStrep. Store at 4 °C and use within two weeks.
Immediately before use, add the antibiotics Nourseothricin (Cat.-No. AB-101) and LEXSY Hygro. (Cat.-No. AB-104) for maintenance of the T7 polymerase and TET repressor genes.

Preparation of glycerol stocks:
Add 1.2 ml of growing culture (ca. 6x10⁷ cells/ml) to one vial with 0.4 ml 80 % glycerol, mix, incubate 10 min at RT, 1 h on ice and over night at -20 °C. Transfer to -80 °C. Strains can be stored this way for several years.

Selected References:
[1] Gruba et al. (2019) Development of Chemical Tools to Monitor Human Kallikrein 13 (KLK13) Activity. Int J Mol Sci. 20: 1557