Team:UCopenhagen/Number-Control

N U M B E R   C O N T R O L

Introduction

Final Design

Experiments

Overview


General verification

Vector creation

  • Biobrick compatible (failed)
  • Vector design
  • CPP tag insertion


Evaluating protein import:

  • Expression of fluorescent proteins
  • Purification and import of the expressed proteins


Verifications and biobrick creation

In all three sub projects, we have used gel electrophoresis and sequencing to verify our stepwise experiments. Read more about our general verifications and biobrick creation under interdependency.

Vector creation

Our vector is a modified version of the pET102 vector, which contains a USER cassette, and a his tag after the USER cassette. The USER cassette is used for cloning genes into. We build the vector design on prSET102; an existing vector in our lab, containing the USER cassette and some restriction sites.

We have made two versions of the vector: one for expression of untagged YFP and BFP, that was also used in the interdependency subproject. The other has a CPP tag before the USER cassette, which will add CPP to the proteins.



Figure 1 Overview of final vectors. First level: The two vectors, unlinearized. Second level: linearized by opening in USER casette. Third level: Insertion of YFP. 4: Produced YFP protein with and without CPP tag. YFP used as example; the same method is used for expression of BFP with and without CPP tag.

Design process/future

References

Find Incell here: