Since our mission is to
create a bacteria
that is
decomposing plastic
, and at the same time adapted to the
harsh
continental climate of Kazakhstan,
we need:
The genes of
Ideonella sakaiensis
bacteria that synthesize the
PETase enzyme.
When we have this gene, we
transform
and
insert this gene into the bacteria
of
the genus Pseudomonas putida, which is able to survive in our climate.
pSB1C3
is a high copy number plasmid (RFC [10])
carrying chloramphenicol resistance.
The replication origin is a pUC19-derived pMB1 (copy number of 100-300 per cell).
pSB1C3 has
terminators bracketing its MCS
which are designed
to prevent transcription from *inside* the MCS from reading out into the vector. The efficiency of
these terminators is known to be < 100%. Ideally we would
construct a future set of terminators
for
bracketing a MCS that were 100% efficient in terminating both into and out of the MCS region.
DesignDesign
Future DesignFuture Design
In the future, our team plans to
continue
working on the project. To do this, we need to carry out a number of mutations on the bacterium P. putida KT2440.
At the moment, the DNA responsible for the synthesis of the PETase enzyme has been
successfully inserted
into the P. putida bacterium, but in the future
we will supplement
it with the genome responsible for the production of the
MHETase enzyme.
Thus, this part was created, which in the future will help us prove our project and create a unique organism with the ability to safely decompose plastic in the southern regions of our country.
