JV4404_delDot.Icm_Lp02

JV4404 is deletion of the Dot.Icm system. Cells were grown in AYE liquid culture with following supplements: ferric nitrate, cysteine hydrochloride and thymidine. Scrape cells off a plate, dissolve in the media, adjust the cell density to 0.6 and make a dilution series. Start O/N culture with all of them and freeze cells next day from the one in early stationary phase that is OD ~ 3.0

JV4404为缺失Dot.Icm系统的菌株。菌体于添加了硝酸铁、半胱氨酸盐酸盐及胸苷的AYE液体培养基中培养。刮取平板上的菌体，将其溶解于培养基内，调整菌体密度至0.6并制备梯度稀释液。将所有梯度稀释物接种并启动过夜培养，次日选取处于早期稳定期、光密度(Optical Density, OD)约为3.0的培养物进行菌体冻存。