Single fragment cloning by Hot Fusion.

(A) A diagram of single fragment cloning in a binary base vector. Base vector containing a lacZ gene was linearized by restriction enzyme digestion with AscI and KpnI to release lacZ. The linearized vector was directly used for Hot Fusion. (B) Amplified PCR products (A1 through A2) of plant gene promoters were used for cloning. M is a NEB 1 kb DNA ladder. (C) Transformation plates of cloned PCR products (A1 and H1 are not shown). Blue colonies on the plates contain the parental vector and white colonies contain the potential recombinants. Eight white colonies were screened for each construct (Table 2).