Part:BBa_K1092003:Experience

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Characterization of BBa_K1092003

1. Biobrick DNA length verification

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

The PCR product meets the target size (936bp), the DNA sequence was further verified by DNA sequencing.

2. Protein expression verification by time series degradation assay

Catechol 1,2-dioxygenase is resposible for phenol degredation (Naiem et al., 2011). Before it is integrated into our PAHs degrdation system, hydroquinone, also known as benzene-1,4-diol or quinol, which is an aromatic organic compound belonging to the phenol family, was used as a substrate to test Catechol 1,2-dioxygenase expression and activity.

Time series analysis were performed. The concentration of hydroquinone was monitored by UV spectrophotometer at 298 nm. According to a chi-square test (Rosenberg et al., 1998), the trends of hydroquinone degradation rate in E. coli BL21 transformed with catechol 1,2-dioxygenase and that in wild type E. coli BL21 were very highly statistically different (p < 0.001) (Figure 2).

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