E. coli strains DH5α and BL21 (DE3), and the plasmid pET-22b and pET-28b are products of Novagen (USA). Human cDNA library was supplied by Maikun Teng, University of Science and Technology of China. The genes encoding the EmGFP, mCherry, the codon-optimized Vhb, the mature mSF (V71-S212) with insertion at BamH I and Xho I sites in the expression vectors, and the helper plasmid for coexpressing yeast mature 5-aminolevulinic acid synthase (ALAS) were constructed by our laboratory [24 (link), 25 (link)].
Heparin Sepharose CL-6B, Phenyl Sepharose CL-4B and amylose resin were obtained from GE (Healthcare, USA). Nickel-nitrilotriacetic acid (Ni–NTA) agarose was purchased from Qiagen (Hilden, Germany). Ultra-15 centrifugal filter tubes equipped with the Ultracel-10 membrane were purchased from Merck-Sigma-Aldrich (Kenilworth, NJ, USA). Primers and the gene encoding the CrLOV were synthesized in General Biol Company (Chuzhou, Anhui, China). Monoclonal antibody including mouse anti-GFP, and anti-MBP, and horseradish peroxidase (HRP) conjugated goat anti-mouse IgG antibody, reagents for Western blot analysis were bought from Transgen Biotech (Beijing, China). RAC was prepared from microcrystalline cellulose, according to our previous report [24 (link)].
Heparin Sepharose CL-6B, Phenyl Sepharose CL-4B and amylose resin were obtained from GE (Healthcare, USA). Nickel-nitrilotriacetic acid (Ni–NTA) agarose was purchased from Qiagen (Hilden, Germany). Ultra-15 centrifugal filter tubes equipped with the Ultracel-10 membrane were purchased from Merck-Sigma-Aldrich (Kenilworth, NJ, USA). Primers and the gene encoding the CrLOV were synthesized in General Biol Company (Chuzhou, Anhui, China). Monoclonal antibody including mouse anti-GFP, and anti-MBP, and horseradish peroxidase (HRP) conjugated goat anti-mouse IgG antibody, reagents for Western blot analysis were bought from Transgen Biotech (Beijing, China). RAC was prepared from microcrystalline cellulose, according to our previous report [24 (link)].