Figure 2

Procedures for generating pBAC-C-KCE-HA base on MAGIC strategy. The donor and recipient plasmids were generated as described in the text, then transformed into the donor strain DH10b and recipient strain DH10B-IS2. The DNA fragments of HA and its expression cassette in the donor vector (pRTHGA-HA), and RFP and its expression cassette in the recipient vector (pBAC-C-KCE) were both cut down by an intron-encoding rare endonuclease I-SceI induced by 0.2% w/v L-arabinose, and then the recombination events were intermediated by the red and gam recombinase induced by 0.2% w/v rhamnose. Then, the recombinant vector pBAC-C-KCE-HA was generated.