Difference between revisions of "Part:BBa K887011"

Latest revision as of 20:14, 3 October 2012

DNA program : specific DNA sequence which can be recognized by zinc fingers

In order to promote the yield of isobutanol further, we not only use the temperature control system we designed but also encode three different kinds of zinc finger functional domain genes (zif268 : Part:BBa_K323105, PBSll : Part:BBa_K323107 and hivC : Part:BBa_K165006) preceding each enzyme genes.(More details about the biobrick, please refer to Part:BBa_K887000)

It means that these fusion proteins (combination of zinc finger functional domain and our enzymes) will bind specific DNA sequence and also carry out isobutanol biosynthesis reaction simultaneously.

The following figure demonstrate specific DNA sequence those three zinc finger functional domains can recognize

If we design a DNA sequence which can make these fusion proteins stay in order, just like a production line, we can achieve our goal, rising yield of isobutanol production because of better efficiency of reaction.
Fortunately, the design of our biobricks has the same order of zinc finger as 2010 Slovenia iGEM team, so we decided to use their DNA program (Part:BBa_K323066) instead of synthesizing one.

However, after sequencing, we found that there is a deletion of a base pair in the zif268 biding site

Thus, we designed a complementary primer to insert the lost base pair. After PCR of point mutation, we sequence it again.
The outcome is below

As you can see, we successfully correct the lost G base at the zif268 binding site and improve the part submitted by 2010 Slovenia iGEM team (Part:BBa_K323066)

Sequence and Features