Team:Michigan/Basic Part

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Basic Parts

BBa_K2105000 (New Part)

The lacZ delta M15 mutation encodes a form of beta-galactosidase lacking residues 11-41. This mutation causes it to be inactive until complemented by the lacZ alpha fragment. The lacZ delta M15 mutant is constitutively expressed in DH5 alpha cells. When the delta M15 mutant and the alpha fragment undergo alpha complementation, they are able to cleave the substrate X-gal to produce a blue product. In our project, the delta M15 mutant is constitutively produced by the cell-free expression system, but the blue output is not produced unless the alpha fragment's transcription is activated in the presence of the target biomarker.

Registry of Standard Biological Parts Entry

BBa_E0033 (New Application of Existing Part)

The lacZ alpha fragment can be used to complement the delta M15 mutant to produce a colorimetric output by cleavage of X-gal. In our system, we use a single stranded version of this biobrick which has been cleaved in the middle of the coding sequence. This prevents transcription of the alpha fragment until a proximity dependent ligation reaction has occurred. Because the rate of this ligation reaction is drastically increased in the presence of a target protein biomarker, transcription of the lacZ alpha fragment only occurs when the biomarker is present.

Registry of Standard Biological Parts Entry