Parts

We submitted three biobrick parts to the parts registry: a composite coding region for the growth factor PDGF-β, an siRNA gene silencer, and an artificial yeast chromosome.

Composite coding region for PDGF-β in yeast (BBa_K635001)

This part contains a platelet derived growth factor (PDGF) sequence modified for effective expression within a yeast chassis. The part contains the cDNA sequence for PDGF codon-optimized for yeast expression. It also contains the biobrick kozak sequence BBa_J63003. Preceding the kozak sequence is a random buffer sequence complementary to an siRNA inhibitor designed for modular inhibition of translation in yeast. The siRNA sequence (BBa_K635000) is reverse complementary to both the kozak sequence and the random buffer. This idea was derived from the 2010 Stanford team's modular siRNA regulation in E.coli. Flanking the cDNA sequence is also a poly-A tail necessary for expression in eukaryotic hosts, such as yeast, to prevent degradation of the mRNA fragment. Following the poly-A tail is the biobrick BBa_K392003 yeast transcriptional terminator. Together these preparatory components allow for expression of PDGF within a yeast chassis.

siRNA gene silencer for use in yeast (BBa_K635000)

This siRNA fragment is designed for modular inhibition of translation for various synthetic biology systems in yeast. The siRNA is reverse complementary to the biobrick kozak sequence BBa_J63003 and a randomly generated buffer sequence that should precede it GGAGTTGTGGGCACCTGCTA. The idea is similar to the RSID system developed by the Stanford 2010 iGEM team for siRNA control in E.coli.

Artificial Yeast Chromosome (Ura3 & Amp) (BBa_K635002)

This part serves as a foundational contribution to expand the registry of yeast components. It is an easy to use artificial yeast chromosome with an auxitrophic uracil selection method for yeast expression, as well as an ampicillin selection for propagation in E.coli. The part was modified for biobrick compatibility from a working pRS316 plasmid containing a Gal1/10 bidirectional promoter which was removed through restriction digest. In addition to the biobrick prefix and suffix, the backbone contains Kpn1 and Sac1 restriction sites the flank the prefix and suffix respectively.

Yeast transformants can be selected by using a uracil drop out medium and E.coli transformants may be selected using ampicillin resistance.