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<i>Third approach</i><br> | <i>Third approach</i><br> | ||
We then decided we could use a pET28a(+) vector of our supervisor that already contained a 14-3-3 dimer. A new gBlock, comprising the third and fourth monomers and GFP, was placed behind our supervisor’s dimer using SacI and HindIII restriction sites and this finally resulted in a successful tetramer with GFP. | We then decided we could use a pET28a(+) vector of our supervisor that already contained a 14-3-3 dimer. A new gBlock, comprising the third and fourth monomers and GFP, was placed behind our supervisor’s dimer using SacI and HindIII restriction sites and this finally resulted in a successful tetramer with GFP. | ||
− | Gel electrophoresis after colony PCR showed that most colonies merely contained the original dimer (around 1700), but two colonies did give good results (around 4200). The sequencing services of StarSEQ confirmed that the formation of a tetrameric 14-3-3 with GFP was finally successful.</h6> | + | Gel electrophoresis after colony PCR showed that most colonies merely contained the original dimer (around 1700), but two colonies did give good results (around 4200). The sequencing services of StarSEQ confirmed that the formation of a tetrameric 14-3-3 with GFP was finally successful.</h6><br/><br/> |
+ | |||
+ | <h6><div class="Figure_3"><img src="https://static.igem.org/mediawiki/2017/4/4c/T--TU-Eindhoven--14-3-3_tetramer_gel.png"> | ||
+ | <figcaption>Figure 1: 14-3-3 tetramer with GFP</figcaption></div></h6> | ||
Revision as of 13:33, 1 November 2017