Team:Shenzhen/Result

Highlights

Project YAO could been seen as pioneering work in iGEM and synthetic biology.

1. When we added some new categories into PartsRegistry, organelles as mitochondria, have opened their access to iGEMers and synthetic biologists.

2. We have taken the initial success, on sending signals in or out organelles, and engineering transcriptional regulation in organelle for the first time.

3. We have been trying to engineer, to measure and to theoretically model a series of signal peptides, related with mitochondria and chloroplasts.

4. We have carried out lots of mathematical simulations, illustrating an exciting blueprint of YAO in bio-synthesis.

YAO.Genome

We have fulfilled the first stage and some work of the second stage, mitochondria expressed GFP in two strategies. We have designed many standardized part for the second stage, shown in table of BioBricks. However, for lime limited, we have not carried out test and standardization. Luckily, we have succeeded in capturing positive clones from the homologues recombination and shuttle plasmid experiments.

YAO.Channel

    We have designed a series of signal peptide BioBricks for mitochondria and TIC & TOC complex from chloroplast.

    Due to some inaccurate information in the iGEM website and our own mistakes, some errors accrued when we attempted to link the reporter gene with signal peptide. 2 bases deletion was found in the plasmid backbone for fusion proteins.

    Point mutation experiments have been designed to overcome this issue.

YAO.Sensor

    Biosensor for redox sensing in mitochondria has been designed. BioBricks have been designed and the experiments are on-going.

    We then modeled the sensor’s functions via the method of modular structured kinetics. In order to model the functions of YAO sensor, we divided the procession into sub parts, and the model’s response to dynamic state are tested.

YAO.Suicider

    We designed all BioBricks needed in this module. We have re-engineered some previous BioBricks, for example, lamda-Holin has been modified from BBb standard to BBa standard, start codon in the 5' ending of GAL promoter has been mutated. Function of Holin has been verified in E. coli.

    Due to the time limit, many wet-lab work is still on-going, so we have focused on the BioBricks modification in wiki pages.

YAO.Factory

    In order to estimate to what extent our YAO system has improved the ability of producing IPP, FPP and GPP in the engineered cell, we construct a model for biochemical reactions of mevalonate pathway in yeast.

    At first we plan to model the reactions at equilibrium states. However, as Gibbs free energy for most of the reactions in mevalonate pathway are unavailable, we decide to model them by Michealis-Menton kinetics at the beginning of the reactions when no products have been accumulated.

    Via our simulation, it is obvious that biosynthesis reactions in YAO could be significantly accelerate.