Team:TecCEM/Composite Part
Composite Parts
Composite Parts
BBa_K2719005: Tenascin Domain V Expression device
Tenascin was modified and optimized for Escherichia coli BL21-DE3 in order to be detected and processed by this strain. In addition, a GTS protein was fusionated to tenascine by a polylinker to conserve the three dimensional structure, because GST provides stability to the Tenascin domain and, also, is a protein that was useful for purification in the case HIS tag doesn’t worked well. This part contain a T7-lac promoter (BBa_R0184), and RBS (BBa_J61100) and a rrnB-T1 terminator (BBa_B0015) using RFC10 assembly standard, thus containing the standard BioBrick prefix and suffix.
BBa_K2719007: Collagen V - like Expression device
Collagen is optimized for Escherichia coli Rosetta 2-DE3 because it was the only strain modified for the detection of non-canonical amino acids. In this case, collagen has a non-canonical amino acid, specifically Hydroxyproline, in order to obtain a correctly binding between collagen and heparin. This protein was also cloned in pSB1C3 and in it, additionally, was cloned a Red Fluorescent Protein (RFP) to prove that the expression of collagen is taking place in the bacteria. This part contain a T7-lac promoter (BBa_R0184), and RBS (BBa_J61100) and a double TAATAA terminator (BBa_B0015). The prefix and suffix of every part were based on BioBrick RFC10.
BBa_K2719009: Leptin expression device
This part is designed for the expression of leptin in E.coli BL21 (DE3), for that function it consists on a T7-lac promoter (BBa_R0184), RBS (BBa_J61100), the leptin coding site (BBa_K2719008) and a double terminator (BBa_B0015). This part was synthetized and the sequence was designed using the standard BioBrick assembly, therefore it has the BioBrick suffix and prefix. This part allows for an inducible expression of recombinant leptin.
BBa_K2719010: Leptin coding device with blue chrome protein
This part facilitates the selection of transformed colonies for the production of leptin (BBa_K2719009) by the incorporation of a reporter cassette (BBa_K592015), which constitutively expresses blue chrome protein. This reporter simplifies the selection of colonies by the appearance of blue colored colonies, serving as transformation and ligation control.
BBa_K2719011: Collagen coding device with blue chromoprotein
This part facilitates the selection of transformed colonies for the production of collagen (BBa_K2719007) by the incorporation of a reporter cassette (BBa_K592015), which constitutively expresses blue chrome protein. This reporter simplifies the selection of colonies by the appearance of blue colored colonies, serving as transformation and ligation control. This part has both reporter genes, RFP (BBa_I13521) and Blue chromoprotein, so it could be easier to detect and select colonies of our interest.
BBa_K2719012: TCD5 coding device with blue chrome protein
This part facilitates the selection of transformed colonies for the production of Tenascin (BBa_K2719005) by the incorporation of a reporter cassette (BBa_K592015), which constitutively expresses blue chrome protein. This reporter simplifies the selection of colonies by the appearance of blue colored colonies, serving as transformation and ligation control.
BBa_K2719013: Blue Chromoprotein and RFP combined reporter
Combined reporter basic model, this part incorporates a constitutively expressed blue chromoprotein cassette (BBa_K592015) and a Tet repressible RFP expression cassette (BBa_I13521). This part serves as a model for the development of multiple reporter systems and how a colony would look when there are multiple reporter proteins and expressed or repressed at the same time. This part improves the understanding of multiple reporter systems and the interactions of two reporter proteins, letting to the understanding of the observations of those systems. Also, was made as a control for collagen (BBa_K2719011) that has the two reporter genes.
