Extended Data Fig. 10: The 2nd entry vector and destination vector construction.
From: Targeted base editing in the plastid genome of Arabidopsis thaliana
a, The 2nd entry vector construction. The 2nd entry vector used in Arimura et al.10, and RECA1 plastid transit peptide coding sequence (PTP; Supplementary Table 4) were amplified with primers corresponding to the DNA template (Supplementary Table 5). The purified PCR products (1: PTP + 2nd entry vector, 2: PTP + destination vector) were mixed with 5× In-Fusion HD Cloning Enzyme Premix (TaKaRa) and incubated at 50 °C for 15 minutes. b, The destination vector construction. The destination vector used in Arimura et al. (2020) was amplified with primers (Supplementary Table 5); purified PCR product was mixed with the PTP PCR product and 5× In-Fusion HD Cloning Enzyme Premix and incubated at 50 °C for 15 minutes to make the destination vector for PTP-GFP expression vector construction. The assembled destination vector was digested by KpnI and the purified product was incubated at 50 °C for 15 minutes after mixed with 5× In-Fusion HD Cloning Enzyme Premix and OLE1-GFP coding sequence amplified from pFAST02 (INPLANTA INNOVATIONS INC), to make the destination vector for ptpTALECD expression vector construction.
