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Genetic modification in
Synechocystis
is done by modifying the chromosome,
rather than inserting plasmids. Synechocystis is naturally transformable, and will
undergo homologous recombination between its chromosome and a plasmid
containing a homologous region. This means that plasmids for insertion or
deletion need to have regions of ~500 to ~1000bp either side of the inserted
sequence, making modifications time consuming or costly depending on your
method of plasmid construction. We will therefore submit all our BioBricks for
insertions and deletions to the registry. All of these will contain Kanamycin
resistance for simple selection of transformants.
Above: Wild-type Synechocystis transformed with a plasmid carrying kanamycin resistance.
The mechanism for each of the BioBricks is the same. They will undergo
recombination with the region that they share homology with. Knockouts will
undergo recombination with the specified gene, replacing it with kanamycin resistance. Insertions will undergo recombination with a region of the
chromosome that is not important for the metabolic
conditions we are using, and will insert the gene of interest along with a kanamycin resistance gene.
We are creating 4 BioBricks that will be submitted
to registry, consisting of 2
deletions (PsaD and PilT1) and 2 insertions (PetF and PilA1). Background information on the aim of these parts can be
found in our project section.
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Reading iGEM 2014 parts |
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<groupparts>iGEM013 Reading</groupparts>
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