Team:GeorgiaState

The 2012 Georgia State University iGEM Team strives to standardize the glyceraldehyde-3-phosphate dehydrogenase (pGAP) promoter shuttle vector to be used in the methylotrophic yeast Pichia pastoris. Creating a standardized expression system within this eukaryotic organism allows the expression of a variety of proteins that can be used for many biological purposes. P. pastoris, in comparison to the conventional recombinant host E. coli, is an ideal choice for the expression of complex proteins due to its ability to perform post-translational modifications and provide a secretion system for these molecules. Modifying the pGAP vector to be used with the iGEM standard enables Georgia State and other teams to utilize P. pastoris as an expression host. In the future, we also plan to standardize the pPic9 shuttle vector to the iGEM standard.

Agrobacterium tumefaciens is a gram-negative soil bacterium and the causative agent of Crown-Gall disease in both dicotyledon and monocotyledon plants. The tumors caused by Crown-Gall disease contain segments from the Tumor Inducing (Ti) Plasmid known as the TDNA, which completely integrate within the plant’s genome. The ability of the Ti plasmid to insert foreign DNA into a plant’s chromosome is used in the field of bio-engineering to manipulate plants to express desired traits. Modification of a gene of interest and insertion into the Ti Plasmid of A. tumefaciens allows exploitation of the natural ability of A. tumefaciens to transmit DNA. In genetic engineering, a binary vector system comprised of only a portion of the Ti plasmid is used because the entire Ti plasmid is over 250 kb, far too large to be easily manipulated. The binary vectors consist of small plasmids with a cloning site and a selectable marker between the left and right border of the TDNA. The aim of our project is to standardize a binary vector system within the parameters of the iGEM competition to be used for the expression of foreign proteins within plants