Naples
From 2007.igem.org
(→Yeast Strain) |
(→Materials & Methods) |
||
| Line 43: | Line 43: | ||
== Materials & Methods == | == Materials & Methods == | ||
| - | We have adopted a strategy of parallel cloning | + | We have adopted a strategy of parallel cloning .Two different reporter genes (luciferase and B-galattosidasy)are cloned in parallel into a vector containing four different promoters. |
* Cloning Strategies | * Cloning Strategies | ||
**Vector choice | **Vector choice | ||
**Restriction Enzyme | **Restriction Enzyme | ||
**Primers Design | **Primers Design | ||
Revision as of 20:20, 18 July 2007
Contents |
University of Naples "Federico II"
Tigem
The Telethon Institute of Genetics and Medicine (TIGEM) was created by the Italian Telethon Foundation in 1994. TIGEM mission is the understanding of the pathogenic mechanisms of genetic diseases with the aim of developing preventive and therapeutic strategies.TIGEM currently hosts 17 research groups, and a total of more than 120 persons, including students, postdoctoral fellows, staff scientists, technicians, and administrators and offers training programs in medical human genetics and Synthetic Biology in cooperation with the University of Naples Federico II.
== About Us == 
- Students:
- Giovanni Russo
- Lucia Marucci
- Velia Siciliano
- Irene Cantone
- Roberta Bergamasco
- Maria Aurelia Ricci
- Mafalda Graziano
- Instructors
- Diego di Bernardo
- Maria Pia Cosma
- Mario di Bernardo
- Advisor
- Giulia Cuccato
Our Project
The aim of our project is to engineer a synthetic biological network and modifying Saccharomyces cerevisiae cells so that we would be able to change colour at differents oleate concentrations.
Oleate is the principal olive oil element and acidity indicator. The olive oil is defined "extra vergine" if it has an acidity lower than 0.8 %,"vergine" with an acidity lower than 2% and not commestible if has an acidity higher than 3%. Oleate induces the transcription of genes involved in peroxisome biogenesis and stimulates the proliferation of these organelles in Saccharomyces cerevisiae. Fatty acid-mediated induction is based on a dramatic increase in transcription of several genes encoding peroxisomal functions due to the presence of an oleate response element (ORE) in their promoters.This upstream activating sequence is minimally defined by an inverted repeat of CGG triplets separated by a 15-18-nucleotide spacer. It constitutes the binding target for the transcription factors Oaf1p and Pip2p.
Yeast Strain
Yeast strain used is W303. All DNA manipulations and subcloning were done in Escherichia coli.
Materials & Methods
We have adopted a strategy of parallel cloning .Two different reporter genes (luciferase and B-galattosidasy)are cloned in parallel into a vector containing four different promoters.
- Cloning Strategies
- Vector choice
- Restriction Enzyme
- Primers Design
