Thermus thermophilus TnSeq
Contents
This site contains the data, scripts and results of a TnSeq experiment performed in Thermus thermophilus HB27 in the laboratory of Mario Mencía & José Berenguer (CBMSO, UAM-CSIC). This analysis was performed by Modesto Redrejo Rodríguez (Biochemistry Department, UAM).
In this study we develop a procedure for thermostable selection of random insertion mutants based on a gene cassette encoding a thermostable resistance to kanamycin flanked by the recognition sites (ME sites) for Tn5 transposase (Kia et al. 2017). The transposition library generated in vitro was initialy transformed in a PrimPol mutant HB27 derivative (ppol), which shows ~2 log fold transformation efficiency (García-Quintans et al. 2020). This allowed the generation of the generation of a large library of insertion mutants (“Mother”), which was later transferred to a wild type HB27 strain (“Daughter”). Both libraries were sequenced straightforward and after several culture passages. See the manuscript for more details.
Funding
This work was supported by grant number PID2019-109073RB-I00 from the Spanish Research Agency (AEI, Spanish Ministry of Science and Innovation).
MRR lab is funded by grant PID2021-123403NB-I00 from the Spanish Research Agency (AEI, Spanish Ministry of Science and Innovation (MCIN/AEI/10.13039/501100011033/FEDER, A way to make Europe).