Team:Freiburg/Parts

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Biobricks

Hier fehlt auf jeden Fall noch die Nennung unseres Favorite Biobricks mit erklärung warum das so ist. Ich denke in unserem Fall wird das der pOP sein oder? (Stefan)
Bitte auf Artnamen achten (Vorname Groß, Nachname klein, alles kursiv (Philipp)) Bezüglich pop: warum ist pSB1C3 ein doofes Backbone für die expression? warum wird iGEM viel mehr proteinexpression machen (in-vitro ist en vogue)? Warum habt ihr plötzlich angefangen ein anderes Backbone genutzt? (Resistenz, lacI etc). Kurz: Warum ist pOP cool?
neuer Versuch (9.9.15 Lara)
Ramona: Hab nochmal etwas umgebaut. Denke, der pOP kommt jetzt besser raus, die Details dazu findet man aber nach wie vor auf seiner eigenen Seite, die jetzt allerdings auch direkt im Text verlinkt ist.

Our project involved the expression of many antigenic peptides. Their immunodominant properties make them feasible for many medical purposes. As ‘Health and Medicine’ is one of the most popular tracks chosen by iGEM teams, we want to share the sequences encoding for these peptides with the iGEM community. Thus, future iGEM teams have the opportunity to take advantage from our research if they are planning to work in the field of diagnostics.
Most of the sequences were obtained by paper research and have been synthesized by Integrated DNA technologies, engaging their special offer for iGEM teams. Before ordering, we used the IDT codon optimization tool to optimize the sequences for efficient expression in E. coli. Planning to submit the sequences to the registry, recognition sites for restriction enzymes used for standard cloning have been removed without causing amino acid changes.
The group of Prof. Dr. Michael Hust (TU Braunschweig) provided us with an antigen specific for Salmonella Typhimurium and a corresponding single chain variable fragment (scFv). They generated the scFv themselves after identifying the immunodominant properties of the protein. Thanks to their work we are able to provide the registry with a specific pair of antigen and scFv.
All our BioBricks are summarized in the table below. Additionally, the table guides you to more detailed info pages as well as to the corresponding registry pages.

Although these parts significantly expand the registry in terms of diagnostics, none of them is our favorite BioBrick. To reach the high yields of protein expression we needed for our project, we used a commercial vector optimized for inducible protein overexpression. This was unavoidable as inserting the required parts (like the lacI gene or a lacI repressible promoter) into pSB1C3 did not result in satisfying protein amounts. Most likely this was due to the high copy origin of replication wasting too much metabolic capacity on replication.
So far, the only plasmid backbone in the Registry with a low-copy origin of replication is pSB6A1. Therefore, we developed an improved version of this plasmid backbone by combining it with a commercial expression vector. Uniting features required for efficient protein overexpression and elements related to iGEM standard cloning, we enable future iGEM teams to perform highly efficient protein overexpression based on our new backbone. Since working in vitro is an upcoming issue in synthetic biology, we decided to chose the backbone as our favorite BioBrick.
Find out more about the pOP vector and see if it also meets your requirements!

BioBrick short description detailed desription
BBa_K1621009 Standardized plasmid backbone optimized for protein overexpressionpOP
BBa_K1621007 scFv binding specifically to the Salmonella Typhimurium derived antigenpRIG15_13
BBa_K1621006 Salmonella Typhimurium specific antigenic protein (DHAD)pRIG15_15
BBa_K1621005 Treponema pallidum specific antigenic peptides derived from bacterioferritin (TpF1)pRIG15_18
BBa_K1621004 Human Immunodeficiency Virus specific antigenic epitopes derived from a polyprotein called gag/tat/pol/envpRIG15_17
BBa_K1621003 Clostridium tetani specific antigenic epitopes derived from tetanus neurotoxin (TeNT_Hc)pRIG15_11
BBA_K1621002 Herpes Simplex specific antigenic epitopes derived from glycoprotein GpRIG15_8
BBa_K1621001 Varicella Zoster Virus specific antigenic epitopes derived from glycoprotein EpRIG15_7
BBa_K1621000 Rubella Virus specific antigenic epitopes derived from glycoprotein E1pRIG15_6