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)
Erster Versuch das Fav-Biobrick etwas genauer zu erklären. Verbesserungsvorschläge willkommen :) (Lara) 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?

Since our project involved expression of many antigenic peptides we decided to share those antigen-sequences with the iGEM community. We obtained most of the sequences via paper research and we would like to give special thanks to the group of Prof. Dr. Michael Hust (TU Braunschweig) who provided us with expression plasmids for the Salmonella Typhimurium antigen and a corresponding single chain variable fragment. The codon optimization tool from Integrated DNA Technologies was used to improve most sequences for expression in E. coli. We also removed all restriction sites that are not allowed in RFC[10], so that all sequences are compatible with the submission vector pSB1C3. When we started working with our first plasmids for this project, we decided to use a specific nomenclature. Every plasmid name starts with “pIG15” which is short for “plasmid igem 2015”. According to this, we named the plasmids containing our biobricks in the shipping backbone pRIG15 (“pRIG” as in “brick”)

In the table below we listed all our biobricks and our biobrick improvement, pOP. We decided on pOP as our favorite biobrick because it provides the iGEM communitiy with an igem conform backbone for protein expression. When we started our project we faced the problem of pSB1C3 being a plasmid designated for cloning colliding with our need of a plasmid for expression of proteins. To be able to express proteins in a vector that exhibits all the properties of an iGEM standard backbone, we improved pSB1C3 (or whatever biobrick we improved) in a way that makes it possible to express proteins in E.coli. Details on which parts of pSB1C3 we changed and how we did this can be found here.

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