Difference between revisions of "Part:BBa K1092003:Experience"
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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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Time series analysis were performed. The concentration of hydroquinone was monitored by UV spectrophotometer at 298 nm. Empty vector-bearing (EV) and dioxygenase-overexpressing cells (T7-Dioxygenase) were induced by 0.3 mM IPTG at OD600 = 0.4, followed by 100 mg/L hydroquinone treatment. Cells were removed and extracellular hydroquinone was determined by OD298 at several intervals. Values represent the average of triplicate samples; error bars denote S.E.M. Paired t-test showed a significant difference in hydroquinone degradation trends between EV and dioxygenase-overexpressing cells (p ≤ 0.05)(Figure 2).
  
  

Revision as of 03:30, 4 October 2013

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Characterization of BBa_K1092003

1. Biobrick DNA length verification

The inserted biobrick DNA length (936 bp) was confirmed by restriction digestion and agarose gel electrophoresis (Figure 1).

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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. Empty vector-bearing (EV) and dioxygenase-overexpressing cells (T7-Dioxygenase) were induced by 0.3 mM IPTG at OD600 = 0.4, followed by 100 mg/L hydroquinone treatment. Cells were removed and extracellular hydroquinone was determined by OD298 at several intervals. Values represent the average of triplicate samples; error bars denote S.E.M. Paired t-test showed a significant difference in hydroquinone degradation trends between EV and dioxygenase-overexpressing cells (p ≤ 0.05)(Figure 2).


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