-Visualization-

-Plasmid extraction

Promoter - suppressed expression system(Am)

-Reining

Above samples

-Make competent cells

F+ and RP4

Cultured to OD600:0.6

Harvested

Suspended with 0.1 M CaCl2

Incubated on ice for 10 min

Harvested and abandoned supernatant

Suspended 0.1 M CaCl2

-Transformation

Promoter - suppressed expression system for F+

oriT - GFP for RP4