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From 2014.igem.org

Contents 1 Introduction 2 NpuDnaE intein RFC [???] circularization constructs 2.1 Design 2.2 Cloning 2.3 Results 3 Sortase A circularization constructs 3.1 Design 3.2 Cloning 3.3 Results? 4 References

Introduction

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