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. 2019 Feb 14;13(1):199–209. doi: 10.1111/1751-7915.13374

Figure 2.

Figure 2

Scheme of markerless recombineering of P. putida using the RecET and Cre vectors. This figure shows more detailed procedures of what we previously described (Choi et al., 2018).
A. Overall procedure of markerless recombineering of P. putida described in subsection ‘Recombineering of P. putida for gene knockout and integration’ of protocols. Bold numbers indicate corresponding steps of the protocol. Refer to the steps in the protocol for detailed description. For steps x to xv, ampicillin may be added to the plates to maintain the RecET vector for subsequent rounds of recombineering. Dashed lines indicate removal of corresponding element from recombinant strains during incubation. Ap, ampicillin; Km, kanamycin; Tc, tetracycline.
B. Vector map of the RecET vector pJB658‐recET. bla, ampicillin resistance gene; RK2, RK2 origin of replication; trfA, gene coding replicase of RK2 origin.
C. Vector map of the Cre vector pRK2Cre. aph(3ʹ)‐Ia, kanamycin resistance gene; trfAts, temperature‐sensitive version of the trfA gene.