Difference between revisions of "Part:BBa K1111007:Experience"

Revision as of 21:34, 30 September 2013

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.

User Reviews

UNIQd2cce9e059677239-partinfo-00000000-QINU UNIQd2cce9e059677239-partinfo-00000001-QINU

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 __NOTOC__
+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.
 ===User Reviews===