Part:BBa_K1111007:Experience

After the plasmid was complete, bacteria were transformed with it and plated. A colony was then picked and the plasmid was sent for sequencing while a glycerol stock of the colony was made. After the sequence was verified, part of the stock was inoculated overnight and then induced by adding 50ul of 20% arabinose to the 5ml liquid culture medium, the expected result being secretion (or at least expression) of the MMP2 protein. A Western blot was performed with anti-His tag antibodies on the super stand and lysate of the bacteria. The result was unfortunately negative. A His-tag purification was then performed and followed by an SDS-PAGE.

Applications of BBa_K1111007

This plasmid can be used for targeted secretion of MMP2 protein. The promoter driving the expression can be replaced by one whose induction conditions suit the user.

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