pCB4270V4BU-amy

Protocol tips

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	The plasmids were purified from E. coli library using GeneAll® Hybrid-QTM Plasmid Rapidprep kit (GeneAll, Seoul, Korea) and transformed into L. citreum CB2567 by electroporation using the following parameters: 25 μF, 1.0 kV, and 400 Ω49. Transformed cells were recovered by incubation on MRS agar plates with 10 mg/L chloramphenicol at 30 °C for 36 h.

Protocol tips
The plasmids were purified from E. coli library using GeneAll® Hybrid-QTM Plasmid Rapidprep kit (GeneAll, Seoul, Korea) and transformed into L. citreum CB2567 by electroporation using the following parameters: 25 μF, 1.0 kV, and 400 Ω49. Transformed cells were recovered by incubation on MRS agar plates with 10 mg/L chloramphenicol at 30 °C for 36 h.

Publication protocol

For the construction of SD2 library, pCB4270B-sfGFP was used as the template DNA, and SD2 region was randomized by PCR with three primers (F2-BCD-et, F1-BL, and R-sfG). The PCR product was digested with XhoI and SalI and ligated into pCB4270B-sfGFP. For the construction of promoter library, pCB4270BU-sfGFP was used as the template, and P710 promoter region was randomized by PCR with two primers (F-P710RL and R-sfG). PCR product was digested with HindIII and SalI, and further was ligated into pCB4270BU-sfGFP. Each library was transformed into E. coli XL1-Blue by electroporation. The plasmids were purified from E. coli library using GeneAll® Hybrid-QTM Plasmid Rapidprep kit (GeneAll, Seoul, Korea) and transformed into L. citreum CB2567 by electroporation using the following parameters: 25 μF, 1.0 kV, and 400 Ω49. Transformed cells were recovered by incubation on MRS agar plates with 10 mg/L chloramphenicol at 30 °C for 36 h.

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