Team:Wisconsin-Madison/Parts

Basic Parts

GSMT is a plasmid gene (854 bp) that codes for the enzyme that catalyze the first reaction in the salt tolerant pathway that we are interested. GSMT stands for Glycine Sarcosine N-Methyl Transferase. It catalyzes the conversion from glycine to sarcosine, using SAM as a methyl donor. In our project, we have this GSMT synthesized using MR. Gene. Naturally, this gene can be found in Galdieria sulphuraria, an extremephile that establishes high resistance to osmotic stress as well as acidic, thermal stresses and toxic metals. We PCR amplified gene GSMT with iGEM designated cutting sites and ribosome binding sites and biobricked it with pSB1A2.

SDMT is a plasmid gene (900 bp) that codes for the enzyme that catalyze the first reaction in the salt tolerant pathway that we are interested. This enzyme of this pathway is SDMT, which stands for Sarcosine Dimethylglycine Methyl Transferase. It catalyzes two steps of methylation, both that of sarcosine and dimethylglycine, to our final product, betaine, again using SAM as the methyl donor. The gene we used came from Galdieria sulphuraria, an extremephile that establishes high resistance to osmotic stress as well as acidic, thermal stresses and toxic metals. In our project, we really appreciate that Center for Eukaryotic Structural Genomics at University of Wisconsin-Madison graciously gave us the plasmid containing this gene. We PCR amplified gene SDMT with iGEM designated cutting sites and ribosome binding sites and biobricked it with pSB1A2.

The ProU promoter (658 bp) was obtained from wild type K12 E. Coli Genomic DNA. Pro U controls transcription of the Pro U operon which contains three sub-units instrumental in bringing choline and proline into the cell. The promoter is most active during times of osmotic stress. We PCR amplified the gene proU with iGEM designated cutting sites and ribosome binding sites and biobricked it with pSB1A2.

NudF is a plasmid gene (560 bp) that codes for the protein responsible for the enzymatic reaction in mevalonent pathway to form Dimethylallyl pyrophosphate (DMAPP). Specifically, in our project, we used it in the last step of mevalonate pathway to produce biofuels iso-pentenol. We PCR amplified gene nudF with iGEM designated cutting sites and ribosome binding sites and biobricked it with pSB1A2.

Composite Parts