Device

Part:BBa_K1092003:Experience

Designed by:   Group: iGEM13_Hong_Kong_CUHK   (2013-09-15)
Revision as of 03:28, 4 October 2013 by MaryZhu (Talk | contribs)

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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. 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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UNIQ369925a6a6706d9f-partinfo-00000000-QINU UNIQ369925a6a6706d9f-partinfo-00000001-QINU