Tn-seq analysis of M1T1 5448 Group A Streptococcus (GAS) genetic fitness under Metal Stress Conditions

To identify genes important for GAS (Streptococcus pyogenes) fitness in the presence the metals Zinc (Zn) and Copper (Cu), an M1T1 5448 master library of Krmit transposon insertions was grown in a defined modified RPMI (mRPMI) media either alone, or in the presence of low (10 µM) and high (20 µM) Zn/Cu for second (T2) or third (T3) overnight passage at 37°C. Transposon-specific Tn-seq insertion tags were generated and sequenced on the Illumina platform prior to fitness analysis.