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Cellular and Molecular Life Sciences

, Volume 68, Issue 7, pp 1207–1214 | Cite as

Technologies of directed protein evolution in vivo

  • Artem Blagodatski
  • Vladimir L. KatanaevEmail author
Review

Abstract

Directed evolution of proteins for improved or modified functionality is an important branch of modern biotechnology. It has traditionally been performed using various in vitro methods, but more recently, methods of in vivo artificial evolution come into play. In this review, we discuss and compare prokaryotic and eukaryotic-based systems of directed protein evolution in vivo, highlighting their benefits and current limitations and focusing on the biotechnological potential of vertebrate immune cells for the generation of protein diversity by means of the immunoglobulin diversification machinery.

Keywords

Directed evolution Protein engineering Mutator strain Cell culture-based systems Somatic hypermutation Gene conversion 

Abbreviations

AID

Activation-induced deaminase

BFP

Blue fluorescent protein

EGFP

Enhanced GFP

FACS

Fluorescence-activated cell sorting

GFP

Green fluorescent protein

GPCR

G protein-coupled receptor

IPTG

Isopropyl β-d-1-thiogalactopyranoside

mRFP

Monomeric red fluorescent protein

PCR

Polymerase chain reaction

TSA

Trichostatin A

Notes

Acknowledgments

The work was supported by grant No. 02.740.11.5016 from the Russian Ministry of Science and by Deutsche Forschungsgemeinschaft (TR SFB-11) to V.L.K. A.B. is a recipient of the Russian state scholarship No. 8084 p/12612 “Participant of the Youth Contest for Science and Innovation”.

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© Springer Basel AG 2010

Authors and Affiliations

  1. 1.Institute of Protein ResearchRussian Academy of SciencesPushchinoRussian Federation
  2. 2.University of KonstanzKonstanzGermany

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