Team:Heidelberg/pages/Circularization Constructs

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Contents

Introduction

The most promising approaches to circularize proteins are protein trans-splicing using split inteins [[#References[1]]] such as Npu DnaE [[#References[2]]] and Sortase A-catalyzed cyclization [[#References[3]]].


On DNA level, creating circular proteins equals creating fusion proteins. However, existing protein fusion standards like RFC 23 cause scars. 


NpuDnaE intein RFC [???] circularization constructs

- Bild: 000, 001

Design

Cloning

mechanismus?

Results

Sortase A circularization constructs

- Bild: 002, 003

Design

Cloning

Results?

References

[1] Iwai, H., Lingel, a & Pluckthun, a. Cyclic green fluorescent protein produced in vivo using an artificially split PI-PfuI intein from Pyrococcus furiosus. J. Biol. Chem. 276, 16548–54 (2001).

[2] Zettler, J., Schütz, V. & Mootz, H. D. The naturally split Npu DnaE intein exhibits an extraordinarily high rate in the protein trans-splicing reaction. FEBS Lett. 583, 909–14 (2009).

[3] Antos, J. M. et al. A straight path to circular proteins. J. Biol. Chem. 284, 16028–36 (2009).