Part:BBa_K887011
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| n/a | We find that the original Slovenia DNA program part a base pair missed, and thus it will make the DNA framshift. |