Team:Queens Canada/ChimeriQ/Parts

ChimeriQ - Parts Page

Parts Page

This is the summary of some of our parts, these parts are a combination of our full flic constructs with different part insertions that have been tested. Our E.coli K12 FliC is our main part that consists of the full flic construct without the D3 domain variable. This part can be used to do part insertions within the sequence.

Featured Parts

This is the complete flagellin coding sequence with biobrick standard cut sites removed, under the promoter R0010 and RBS B0034. This part can be used to over express the flagellin monomer.

The main use of this part is for PCR overlap extension, which will make an insertion at the annotated site, given that the insert is in the appropriate format. The resulting chimera will express protein immediately after the PCR reaction is transformed.

This part has been previously characterized as BBa_K777109. The part that we have submitted has some slight differences in the silent mutations made to remove Biobrick cut sites. This part also includes the promoter R0010 and the RBS B0034.

This is the modified RFP protein obtained from J04450, that has been incorporated into the E. coli flagellin as an insertion.

This part was successfully characterized and determined to show expression of the red fluorescent protein from J04500. As can be seen below, there appears to be less detectable fluorescence in cell cultures expressing the different types of chimeras. This result can be expected for a number of possible reasons.

The chimeric protein may not be as stable as the free RFP.

The RFP may be polymerizing in flagella, resulting in a quenching effect as the fluorescent light may be absorbed by other RFP molecules arranged in close proximity.

The flagellin domains may hinder proper folding of the RFP. This may be why the more constrained deletion variant is showing less expression compared to the insertion variant, which has more flexibility.

This is the modified RFP protein obtained from J04450, that has been incorporated into the E. coli flagellin as a deletion.

This part can be used as a template for making other PCR overlap insertions. After transforming the product, and digesting the template plasmid with Dpn1, any template that has not been successfully digested will form red colonies.

This part was successfully characterized and determined to show expression of the red fluorescent protein from J04500. As can be seen below, there appears to be less detectable fluorescence in cell cultures expressing the different types of chimeras. This result can be expected for a number of possible reasons.

The chimeric protein may not be as stable as the free RFP.

The RFP may be polymerizing in flagella, resulting in a quenching effect as the fluorescent light may be absorbed by other RFP molecules arranged in close proximity.

The flagellin domains may hinder proper folding of the RFP. This may be why the more constrained deletion variant is showing less expression compared to the insertion variant, which has more flexibility.

Submitted Parts

This is an mgfp-5 chimeric insertion flagella. mgfp-5 is a mussel foot binding protein.

This is type II cohesin from Clostridium Thermocellum as a chimeric insertion into flagellin.

Rv2579 is a dehalogenase enzyme. This construct as incorporated that enzyme as an insertion into the variable domain.

LinB is a dehalogenase enzyme. It degrades haloalkanes which are toxic pollutants that accumulate in food chains. This part has the LinB incorporated at an insertion site.

This is an XylE deletion variant of the flagellin monomer. The enzyme XylE that degrades catechol was incorporated into the E. coli flagellin by replacing the variable domain.