Development of the Multiple Gene Knockout System with One-Step PCR in Thermoacidophilic Crenarchaeon Sulfolobus acidocaldarius

Figure 2

Effects of recovery conditions on transformation efficiency. SK-1 cells harvested at early to midlog phase (OD₆₀₀ = 0.308) were transformed with 200 ng of pyrElacS800. After electroporation (15 kV/cm, 9 ms), the cell suspension was transferred to 800 μL of recovery buffer (20 mM sucrose, 2× basal4, Buffer C, and MBS) and incubated. Ten percent of the regenerated sample was mixed with 10 mL of top gel solution and poured onto XT plates by overlay cultivation. Error bars represent the standard deviation of three independent experiments.