Difference between revisions of "Part:BBa K2319000"

Revision as of 10:15, 1 November 2017

sfGFP-SpyCatcher under T7 expression system

This part generates a fusion protein of superfolder GFP (sfGFP) and SpyCatcher (a 15.2 kDa protein) when transformed into a T7 expression system like E. coli strain BL21 (DE3).

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This part can be used as a T7 expression backbone.

Using the T7 expression backbone

Expressing any protein of interest

This T7 expression backbone can be used to express any protein if its coding sequence (with a start codon) is inserted using the HindIII and NheI sites. These sites can be added to the coding sequence using PCR with primers having 5'-overhangs.

Fusing a protein domain at the N-terminus of an existing protein

By inserting the coding sequence of a protein domain (including the start codon) using the HindIII and AgeI sites into the T7 expression backbone (which already contains a protein coding sequence), an N-terminal fusion can be performed.

Sequence and Features