Production trial of PHAs by past teams

We introduce some attempts in the past iGEM to show how great our work is in iGEM.

iGEM10_Caltech

They couldn’t prove that their engineered bacteria produced PHB according to the team wiki.

iGEM10_INSA-Lyon

They registered and submitted a part, which contains only phaC sequence, as worked. But the team wiki shows that the part doesn’t meet the standard which is required in iGEM because they couldn’t cut their part with Xba1.

iGEM09_Duke

State iGEM08_Utah State

iGEM08_Hawaii

iGEM08_Virginia

Construction of pha-C1-A-B1 in Biobrick format

fig3,construction of phaC1-A-B1

To construct a part that meets Biobrick format, we have modified the phaC1-A-B1 operon not to contain forbidden restriction enzyme sites. First, we cloned the wild type gene phaC1-A-B1 from R.eutropha H16 by using PCR and inserted the gene into pSB1C3. However, wild type phaC1-A-B1 gene sequence contains one NotI and three PstI recognition sites that are not allowed in Biobrick format. To get phaC1-A-B1 sequence without these recognition sites, we ordered the chemically synthesized DNA from IDT/MBL. In this chemically synthesized DNA, coding is optimized for E.coli. We used restriction enzyme XbaI (on pSB1C3) and BsrGI (on phaC1-A-B1) to insert sequence. That is to say, we got Poly[(R)-3-hydroxybutyrate] synthesizing gene in Biobrick format ([http://partsregistry.org/wiki/index.php?title=Part:BBa_K934001 BBa_K934001]). Fig3:construction of phaC1-A-B1

Production of PHAs by E.coli