Team:Kent/Basic

Basic Parts


We have created eleven basic parts:



BBa_K1985000 pSB1C3-MamP

Figure 1. Shows part BBa_1985000 in pSB1C3. The image was created using Snapgene. The mamP gene from the magnetosome operon mamAB forms a protein dimer. It is proposed to be an iron-oxidase that contributes to the formation of magnetite by mediating one step in the process, the formation of iron(III)ferrihydrite [1]. This biobrick encodes for the wild type protein sequence of mamP from Magnetospirillum gryphiswaldense.



BBa_K1985001 pSB1C3-MamT

Figure 2. Shows part BBa_1985001 in pSB1C3. The image was created using Snapgene. The mamT gene produces an integral membrane protein that can be used to transfer electrons to iron molecules, thereby forming magnetite from iron [2]. This biobrick encodes for the wild type protein sequence of mamT from Magnetospirillum gryphiswaldense.



BBa_K1985002 pSB1C3-MamX

Figure 3. Shows part BBa_1985002 in pSB1C3. The image was created using Snapgene. The mamX gene also produces an integral membrane protein that can be used to transfer electrons to iron molecules, thereby forming magnetite from iron [2]. This biobrick encodes for the wild type protein sequence of mamX from Magnetospirillum gryphiswaldense.



BBa_K1985006 pSB1C3-MamO

Figure 4. Shows part BBa_1985002 in pSB1C3. The image was created using Snapgene. The mamO gene produces a protein that can be used to initiate the formation of magnetite by "nucleating" the crystal particles, allowing further development [2]. The biobrick encodes for the wild type protein sequence of mamO from Magnetospirillum gryphiswaldense.



BBa_K1985003 pSB1C3-[SecS-sol-MamP]

Figure 5. Shows part BBa_1985003 in pSB1C3. The image was created using Snapgene. This biobrick is a variation on part BBa_1985000. It differs from the wild type sequence in that it has a his-tag added, the membrane anchor cleaved and is targeted to the periplasm instead of the membrane.



BBa_K1985004 pSB1C3-[SecS-sol-MamT]

Figure 6. Shows part BBa_1985004 in pSB1C3. The image was created using Snapgene. This biobrick is a variation on part BBa_1985001. It differs from the wild type sequence in that it has a his-tag added, the membrane anchor cleaved and is targeted to the periplasm instead of the membrane.



BBa_K1985005 pSB1C3-[SecS-sol-MamX]

Figure 7. Shows part BBa_1985006 in pSB1C3. The image was created using Snapgene. This biobrick is a variation on part BBa_1985002. It differs from the wild type sequence in that it has a his-tag added, the membrane anchor cleaved and is targeted to the periplasm instead of the membrane.



BBa_K1985016 pSB1A3-[AraC-pBAD]

Figure 8. Shows part BBa_1985016 in pSB1C3. The image was created using Snapgene. BBa_K1985016 is a regulatory part is made up of the arabinose-Inducible promoter, pBAD, and its transcriptional inhibitor/activator, AraC. In the absence of arabinose, AraC inhibits gene expression by binding to the operators present within the AraC and pBAD promoter sequences, and producing a DNA loop that prevents the RNA polymerase and CAP from binding and initiating transcription. When supplied, arabinose binds to the AraC dimer, releasing it from the operator in the pBAD promoter sequence. It also enhances transcription by allowing binding to the enhancer sites upstream [3]. This is a twin of part BBa_K1321333 created by team Imperial in 2014. This part is an improvement as it was constructed in pSB1A3 so that it could be used for co-transformation with another chloramphenicol resistant plasmid or into a chloramphenicol resistant E. coli strain.



Bba_K1985012 pSB1C3-[CsgA-SS-Sup35NM]

Figure 9. Shows part BBa_1985012 in pSB1C3. The image was created using Snapgene. This part is an improved version of a previously designed BioBrick (Part:BBa_K1739002), which was designed by the Kent 2015 iGEM team. The promoter BBa_J23104 has been removed to enable the use of this part using different promotors. The part contains two segments in one biobrick: CsgA signal sequence and Sup35NM.



Bba_K1985010 pSB1C3-[CsgA-SS-Sup35-1-61]

Figure 10. Shows part BBa_1985010 in pSB1C3. The image was created using Snapgene. This part is an improved version of a previously designed BioBrick (Part:BBa_K1739002), which was designed by the Kent 2015 iGEM team. This part contains two segments, the CsgA signal sequence and the first 61 residues of the prion forming domain Sup35. Our improved BioBrick aims to optimize the self-assembly process of amyloid fibrils with the addition of these residues as they have been considered to be a suitable building block for the assembly of functional nanostructures[4].



Bba_K1985011 pSB1C3-[CsgA-SS-Sup35-1-61-Cytb562]

Figure 11. Shows part BBa_1985011 in pSB1C3. The image was created using Snapgene. This part is an improved version of the Envirowire fusion protein(BBa_K1739003). It contains 3 segments, the CsgA signal sequence, the first 61 residues of the prion forming domain Sup35 and the electron transfer protein cytochrome 562.



References
  1. Siponen, Marina I. et al. "Structural Insight Into Magnetochrome-Mediated Magnetite Biomineralization". Nature 502.7473 (2013): 681-684.
  2. Nudelman, Hila and Raz Zarivach. "Structure Prediction Of Magnetosome-Associated Proteins". Front. Microbiol. 5 (2014): n. pag. Web. 17 Oct. 2016.
  3. Artem Khlebnikov, J. D. Keasling. "Regulatable Arabinose-Inducible Gene Expression System With Consistent Control In All Cells Of A Culture". Journal of Bacteriology 182.24 (2000): 7029.
  4. Men, D., Zhou, J., Li, W., Leng, Y., Chen, X., Tao, S., and Zhang, X. "Fluorescent protein nanowire-mediated protein microarrays for multiplexed and highly sensitive pathogen detection". ACS Appl. Mater. Interfaces (June 2016) DOI: 10.102.




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