Dna restriction enzyme

Escherichia coli strains BL21 (DE3) and DH5α from TaKaRa (Dalian, China) were cultivated in Luria–Bertani (LB) medium containing Kanamycin (50 μg/mL) when necessary. For the expression of recombinant proteins, plasmid pET-24a (+) was used. The DNA polymerase and DNA Restriction enzyme were from TaKaRa (Dalian, China). TIANamp Bacteria DNA Kit was purchased from TIANGEN BIOTECH (Beijing, China). ClonExpress II One Step Cloning Kit was purchased from Vazyme (Nanjing, China). Qingdao Gather Great Ocean Algae Industry Group Co., Ltd. (Qingdao, China) provided Alginate and polyM, polyG were from Qingdao HEHAI Biotech Co., Ltd. (Qingdao, China).

Analytical DOP standards were purchased from Dr. Ehrenstorfer GmbH (Augsburg, Germany). The hybridoma cell line (12C2), the coating hapten H1 (4-((diethoxyphosphorothioyl) amino) butanoic acid), and the coating antigen (hapten 2-ovalbumin) were self-prepared [11 (link)], as previously described. The XL1-Blue and Top 10F’ Escherichia coli strains have previously been established by our laboratory. The plasmid vector pComb3XSS (Figure 1A) was obtained from the Barbas Laboratory, TSRI, La Jolla, CA, USA. The helper phage VCSM13 was obtained from the Naval General Hospital of Beijing, China. Horseradish peroxidase (HRP), and 3,3′,5,5′-tetramethylbenzidine (TMB) were obtained from Sigma-Aldrich (Shanghai, China). Ampicillin, kanamycin, and Isopropyl β-d-thiogalactopyranoside (IPTG) were purchased from Takara (Dalian, China). DNA polymerase and DNA restriction enzyme were also purchased from Takara. HRP-conjugated goat anti-mouse IgG and anti-His tag mouse monoclonal antibodies were purchased from TransGen Biotech Co. Ltd (Beijing, China). Mut Express II Fast Mutagenesis Kit V2 for site-specific mutagenesis was obtained from Vazyme Biotech Co., Ltd. (Nanjing, China). All other chemicals were standard commercial analytical-grade reagents.