Part:BBa_P0451:Experience

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User Reviews

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• HZAU-China 2014

We met some problem in our construction. It seems that the overexpression of repressor protein has cytotoxicity which made troubles in our process. We just ligated the c1 protein coding sequence with lac promoter, and repeated for many times. However, the sequencing result suggested that there was always either a deletion of the promoter or an unknown insertion in c1 coding sequence in our samples. We speculate that c1 protein has cytotoxicity and burden the E.coli, so the samples were always with mutations that cause of no expression of c1 protein or its inactivation. When we ligated it with the lac promoter which is repressed by LacI, the ligation is successful. The similar situation also happened on TetR.

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