A nitrite electrochemical sensor based on electrodeposition of zirconium dioxide nanoparticles on reduced graphene oxide modified electrode was successfully constructed for the detection of nitrite. The electrochemical behavior of the modified electrode was investigated by cyclic voltammetry and amperometric i-t curve. Under the optimal conditions, the amperometric i-t curve response of the electrode showed a linear relationship with nitrite concentration in the range of 3.0×10Symbolm_7-1.0×10Symbolm_6 mol/L and 1.0×10Symbolm_6-6.0×10Symbolm_6 mol/L, and the detection limit was 1.0×10Symbolm_7 mol/L (S/N=3). The fabricated sensor exhibited high sensitivity, good stability and high reproducibility. This sensor was applied for the detection of nitrite in sausage samples with favorable recoveries of 93.7％-110.4％ and relative standard deviation (RSD) of 1.6％-2.1％.

To use the designed restriction enzyme assisted mutagenesis technique to perform rapid site-directed mutagenesis on double-stranded plasmid DNA. The target amino acid sequence was reversely translated into DNA sequences with degenerate codons, resulting in large amount of silently mutated sequences containing various restriction endonucleases (REs). Certain mutated sequence with an appropriate RE was selected as the target DNA sequence for designing mutation primers. The full-length plasmid DNA was amplified with high-fidelity Phusion DNA polymerase and the amplified product was 5' phosphorylated by T4 polynucleotide kinase and then self-ligated. After transformation into an E. coli host the transformants were rapidly screened by cutting with the designed RE. With this strategy we successfully performed the site-directed mutagenesis on an 8 kb plasmid pcDNA3.1-pIgR and recovered the wild-type amino acid sequence of human polymeric immunoglobulin receptor (pIgR). A novel site-directed mutagenesis strategy based on DREAM was developed which exploited RE as a rapid screening measure. The highly efficient, high-fidelity Phusion DNA polymerase was applied to ensure the efficient and faithful amplification of the full-length sequence of a plasmid of up to 8 kb. This rapid mutagenesis strategy avoids using any commercial site-directed mutagenesis kits, special host strains or isotopes.
Amino Acid Sequence ; Base Sequence ; DNA ; genetics ; DNA Restriction Enzymes ; genetics ; DNA-Directed DNA Polymerase ; genetics ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; methods ; Plasmids ; Receptors, Polymeric Immunoglobulin ; genetics