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-	{{Template:Tuebingen2012navigation}}	+	{{:Team:Tuebingen/Template/Tuebingen}}
-	__NOTOC__	+
-	*<span style="color:red">This is our provisional page; the final one is still in work. </span>	+
		+	== The Project ==
-	== '''Overview''' ==	+	* [[Team:Tuebingen/ProjectOverview|Overview]] <br /> A short introduction including an animation film.
-	Our general aim is to establish a simple mechanism which allows us to measure an estrogen concentration in a solution as precisely as possible. Therefore we want to clone the membrane progestin receptor (mPR) of the zebrafish (''Danio rerio'') in yeast (''saccharomyces cerevisiae'') and link it to a reporter gene, which allows a quantitative  measurement.	+	* [[Team:Tuebingen/ProjectQuestions|Questions]] <br /> Every project should start with important questions, here are ours.
-		+	* [[Team:Tuebingen/ProjectMechanism|Mechanism]] <br /> An in-depth explanation of our genetically engineered system.
-	== '''Motivation''' ==	+	* [[Team:Tuebingen/ProjectImplementation|Implementation]] <br /> Assembly and plasmid construction to implement the system.
-	Why do we want to establish a mechanism for steroid measurement?	+	* [[Team:Tuebingen/Parts|Submitted Parts]] <br /> All of our submitted Parts.
-		+	* [[Team:Tuebingen/References|References]] <br /> Scientific literature used to design our project.
-	Steroid hormones, especially estrogens, occur in all vertebrates and play a crucial role in sexual differentiation.  In recent times the pollution of waters with these hormones has become an increasing problem for the aquatic fauna.	+
-	Particularly waters functionalized by humans or adjacent to human settlements, for example in areas with agricultural use, show increased estrogen-concentrations.	+
-		+
-	Scientific studies based on ''Danio rerio'' showed that the consequences are devastating.  High concentrations of 17α-ethinylestradiol, a hormone in most  birth-control pills, affected the sex differentiation of ''Danio rerio'' leading to development of ovotestis or complete feminization (Andersenc, 2002).	+
-	Intersex-fish have been reported in UK rivers since 1978 downstream of an sewage treatment plant.	+
-		+
-	We believe that a first step in finding a solution to this environmental problem is an accurate and reliable method to quantify steroids which is easy to use and limited to an optical measurement.	+
-		+
-		+
-	== ''Occurring Questions'' ==	+
-	On our way designing the major pathway to express a specific reporter gene to demonstrate the presence of steroid hormones, we had and still have to deal with several questions concering the choice of BioBricks, genes and vectors to construct a firm method to determine "pollution" by steroid hormones. As a conclusion, we have to meet two major requirements for our system:	+
-	* It should be as cost-efficient as possible for easy and regular application	+
-	* It should be resistant to yeast's own metabolism. (Not be disturbed by unexpected occuring expression)	+
-		+
-	At first we had to find an appropriate receptor to detect steroid hormones.	+
-		+
-	== '''Mechanism''' ==	+
-	[[File:Mechanism_steroid_good_version.jpg]]	+
-		+
-	=== '''Receptors''' ===	+
-		+
-	Membrangebunden, 7-Transmembranrezeptor (Innen C-Terminus, außen N-Terminus), PAQR-Familie (progesterone adiponectin Q receptor), Hly-III superfamily	+
-	G-Protein gekoppelt: Aktivierung von inhibitorische Gi-Einheiten: Verringerung der Aktivität der Adenylyl-Cyclase	+
-	G Proteins (Protein Data Bank) Artikel zu G-Proteinen - Struktur und Funktionsweise, schöne Bilder	+
-		+
-	Sequenz mPR alpha (Zebrafisch): [http://www.ncbi.nlm.nih.gov/nuccore/AY149121.1 NCBI],	+
-		+
-	Sequenz mPR beta (Zebrafisch):  [http://www.ncbi.nlm.nih.gov/nuccore/AY149120.1 NCBI]	+
-		+
-	Homologe zum mPR Alpha Danio rerio:	+
-	*Goldfisch, Carassius auratus	+
-	*Katzenwels, Ictalurus punctatus	+
-	*Hundszungen, Cynoglossus semilaevis	+
-	*Flunder, Paralichthys lethostigma	+
-	*Meerforelle, Cynoscion nebulosus	+
-	*Buntbarsch, Oreochromis niloticus	+
-	*Umberfisch, Micropogonias undulatus	+
-		+
-	Zebrafrisch (Danio rerio)	+
-	In Tübingen:	+
-	*Tübingen Map of the [http://wwwmap.tuebingen.mpg.de/ Zebrafish Genome]	+
-	*[http://www.mnf.uni-tuebingen.de/fachbereiche/biologie/institute/evolutionecology/lehrbereiche/physiologische-oekologie-der-tiere/staff/volker-scheil.html Dr. Volker Scheil], The impact of potential environmental stressors on early development and cellular and biochemical biomarkers in fish, Main research: Fish embryotoxicity, histopathology and stress protein (hsp 70) responses.	+
-		+
-	=== '''Inhibitor and promotor''' ===	+
-		+
-	An appropriate inhibitor/promotor combination is a crucial step in our pathway and should be selected wisely.	+
-		+
-	=== '''Reporter gene''' ===	+
-		+
-	The last station of our signaling pathway should be a reporter gene which amplifies our initial signal to allow a quantitative  measurement.	+
-		+
-	The enzyme '''luciferase''' fulfills these conditions and is possible candidate for our mechanism.	+
-		+
-	== '''Measurement''' ==	+
-		+
-	The measurement itself should be limited to an optical one and people with lesser qualifications should be able to run it.	+
-		+
-	== '''References''' ==	+
-		+
-	*Sumpter, Johnson (2008) Reflections on endocrine disruption in the aquatic environment	+
-	*Liew et. al. - 2012 Polygenic Sex Determination System in Zebrafish	+
-	*Hanna et al. - 2006 - Cell-surface expression, progestin binding, and rapid nongenomic signaling of zebrafish membrane progestin receptors alpha and beta in transfected cells	+
-	*Reupke - 2011 - Detektion dopingrelevanter anaboler Steroide in Pferdeurin und Pferdeplasma mithilfe eines Reportergen-Assays in Hefezellen. (Detection of doping relevant anabolic steroids in horse urin and blood plasma using yeast reporter gene assays)	+
-	*Jessica L. Smith, Brian R. Kupchak, Ibon Garitaonandia, L. Kim Hoang, Andrew S. Maina, Lisa M. Regalla, and Thomas J. Lyons  - 2008 - Heterologous expression of human mPRα, mPRβ and mPRγ in yeast confirms their ability to function as membrane progesterone receptors	+
-	*“Assault on the male”, BBC 1996. Video report http://www.youtube.com/watch?v=LkxIJJI37bQ	+
-	*Dr. Volker Scheil, The impact of potential environmental stressors on early development and cellular and biochemical biomarkers in fish, Main research: Fish embryotoxicity, histopathology and stress protein (hsp 70) responses.	+
-	*Harris et al. - 2011 - The consequences of feminization in breeding groups of wild fish	+
-	*Liew et al. - 2012 - Polygenic Sex Determination System in Zebrafish	+
-	*Sumpter, Johnson - 2008 - 10th Anniversary Perspective Reflections on endocrine disruption in the aquatic environment from known knowns to unknown unknowns (and many things in between)	+
-	*Werner, Palace, Wautier - 2006 - Reproductive fitness of lake trout (Salvelinus namaycush) exposed to environmentally relevant concentrations of the potent estrogen ethynylestradiol (EE2) in a whole lake exposure experiment	+
-	*Fenske, M., Maack, G., Schäfers, C., & Segner, H. (2005). An environmentally relevant concentration of estrogen induces arrest of male gonad development in zebrafish, Danio rerio. Environmental toxicology and chemistry / SETAC, 24(5), 1088-98. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/16110986 (kein PDF)	+
-	*Schäfers, C., Teigeler, M., Wenzel, A., Maack, G., Fenske, M., & Segner, H. (2007). Concentration- and time-dependent effects of the synthetic estrogen, 17alpha-ethinylestradiol, on reproductive capabilities of the zebrafish, Danio rerio. Journal of toxicology and environmental health. Part A, 70(9), 768-79. doi:10.1080/15287390701236470	+
-	*Segner, H., Caroll, K., Fenske, M., Janssen, C. R., Maack, G., Pascoe, D., Schäfers, C., et al. (2003). Identification of endocrine-disrupting effects in aquatic vertebrates and invertebrates: report from the European IDEA project. Ecotoxicology and environmental safety, 54(3), 302-14. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/12651186 (kein PDF)	+
-	*Hanna et al. - 2006 - Cell-surface expression, progestin binding, and rapid nongenomic signaling of zebrafish membrane progestin receptors alpha and beta in transfected cells	+
-	*Thomas et al. - 2007 - Steroid and G protein binding characteristics of the seatrout and human progestin membrane receptor alpha subtypes and their evolutionary origins	+
-	*Thomas - 2008 - Characteristics of membrane progestin receptor alpha (mPR ) and progesterone membrane receptor component 1 (PGMRC1) and their roles in mediating rapid progestin actions	+
-	*Jessica L. Smith, Brian R. Kupchak, Ibon Garitaonandia, L. Kim Hoang, Andrew S. Maina, Lisa M. Regalla, and Thomas J. *Lyons  - 2008 - Heterologous expression of human mPRα, mPRβ and mPRγ in yeast confirms their ability to function as membrane progesterone receptors	+

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Latest revision as of 14:46, 18 September 2012

The Project

• Overview
  A short introduction including an animation film.
• Questions
  Every project should start with important questions, here are ours.
• Mechanism
  An in-depth explanation of our genetically engineered system.
• Implementation
  Assembly and plasmid construction to implement the system.
• Submitted Parts
  All of our submitted Parts.
• References
  Scientific literature used to design our project.