Difference between revisions of "Part:BBa K3187017"

coat protein

Usage and Biology

Coat Protein is an umbrella term for many different proteins, which simplify the transfer of molecules between different compartments that are surrounded by a membrane. ^[1] We focused on the viral and bacteriophagic coat proteins, which are parts of their respective organisms’ capsid. The genetic information (DNA or RNA) is wrapped and protected by the capsid. During cell infection, the phage or virus transfer the genetic information into the infected cell. Because of the high variety of coat proteins, we are focussing on one specific coat protein (BBa_K3187017), which is naturally found in the bacteriophage P22. This coat protein (CP) consists of 431 amino acids and its molecular weight is 46.9 kDa. Because of its significance as a part of the capsid, it represents one main part of our Virus-like particle. Together with the scaffold protein (BBa_K3187021), they assemble to a VLP and form the basis for our modular platform.

Results

The basic part coat portein was expressed, produced and purified as the composite part BBa_K3187000 (coat protein with LPETGG) and BBa_K3187001 (coat protein). The production is performed in E. coliBL21 and it is purified with GE Healthcare ÄKTA Pure machine LINK which is a machine for FPLC. To verify the right production, a Western blot is made.

Figure 1: Western blot of all produced and purified proteins.

Fig. 1 shows that the band of the CP-LPETGG is approximately found by the 49 kDa band and the band of CP by 46.9 kDa. Consequently, the successful cloning and expression was proven. CP-LPETGG and CP are detected with Strep-Tactin-HRP.

References

1. ↑ Juan S. Bonifacino, Jennifer Lippincott-Schwartz, Coat proteins: shaping membranetransport, NATURE REVIEWS MOLECULAR CELLBIOLOGY, May 2013, 4, 409-414 [1]