Fig. 1.

Structural design and construction of 3′ end PCC fusion subunits variants. (a) 3′ end fusion subunits variants were designed to be amplified from original PCC subunits clone. MTS-TAT fusion subunits could be amplified from MTS fusion subunits. (b) A schematic representation of TAT, MTS, and TAT-MTS fusion 5′ end subunits constructs. Control variants are subunits lacking the MTS or TAT domains. (c) PCR amplification results of pccA and pccB variants by using KOD DNA polymerase. (d) Digestion of cloned 3′ end fusion pccA and pccB subunits in pET28a vector by restriction enzymes. pccA-TAT, pccA-MTS and pccA-MTS-TAT clones were digested by EcoRI-HF and HindIII-HF restriction enzymes. pccB-TAT, pccB-MTS and pccB-MTS-TAT clones were digested by NotI-HF and BamHI-HF restriction enzymes. All digestions were carried out in 37 °C overnight and digestion results visualized by GelRed staining on 10% agarose gel.