Part:BBa_K1092005:Experience

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Characterization of BBa_K1092005

1. Biobrick DNA length verification

To check the inserted biobrick DNA length, PCR was conducted (Figure 1).

The PCR product meets the target size (1542 bp), the DNA sequence was further verified by DNA sequencing.

2. Protein activity test by automatic kinetic spectrophotometric assay using o-phenylenediamine

Laccase is capable of oxidizing o-phenylenediamine (1,2-diaminobenzene) during the stopped-flow period. During oxidation, 2,3-diaminophenazin, that can be determined spectrophotometrically, will be produced. So the kinetic spectrophotometric method can be used to detect laccase activity (Huang et al., 1996).

After 10 to 12 hours of incubation at 37 °C, laccase showed maximum activity when 0.3 mM IPTG was used for introducing protein expression (Figure 2).

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