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Cell-Free Systems: Functional Modules for Synthetic and Chemical Biology

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Fundamentals and Application of New Bioproduction Systems

Part of the book series: Advances in Biochemical Engineering/Biotechnology ((ABE,volume 137))

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Abbreviations

ATP:

Adenosine-5’-triphosphate

AzPhe:

p-azido-l-phenylalanine

AzPheRS:

Orthogonal mutant synthetase specific for AzPhe

BP:

Bandpass filter

CECF:

Continuous-exchange cell-free

CFCF:

Continuous-flow cell-free

CHO:

Chinese hamster ovary

CLSM:

Confocal laser scanning microscopy

CrPV:

Cricket paralysis virus

DDM:

n-dodecyl-ß-maltoside

DTT:

Dithiothreitol

E. coli :

Escherichia coli

EndoH:

Endoglycosidase H

E-PCR:

Expression-PCR

EPO:

Erythropoietin

ER:

Endoplasmic reticulum

eYFP:

Enhanced yellow fluorescent protein

Fab:

Fragment, antigen-binding

FITC:

Fluorescein isothiocyanate

Fv:

Antibody variable fragment

GTP:

Guanosine-5’-triphosphate

GUV:

Giant unilamellar vesicle

IGR:

Intergenic region

IRES:

Internal ribosome entry site

KD:

Association equilibrium constant

Mel:

Melittin

PAGE:

Polyacrylamide gel electrophoresis

PDI:

Protein disulfide isomerase

PI:

Protease inhibitors

PNGaseF:

N-glycosidase F

PTM:

Posttranslational modification

RU:

Resonance units

scFv:

Single-chain antibody fragment

SDS:

Sodium dodecyl sulfate

Sf21:

Spodoptera frugiperda 21

SN1:

Supernatant 1

SN2:

Supernatant 2

SPR:

Surface plasmon resonance

TCA:

Trichloroacetic acid

TM:

Translation mixture

tRNACUA :

Amber suppressor tRNA

VF1:

Vesicular fraction 1

VF2:

Vesicular fraction 2

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Acknowledgments

The authors gratefully acknowledge Dipl.-Ing. Doreen Wüstenhagen, Dipl. Nutritional Scientist Conny Mascher, and Birgit Hollmann (Fraunhofer IBMT, Potsdam-Golm, Germany) for preparing cell-free extracts and in general, for keeping things running in our cell-free lab. Furthermore, we thank Dr. Walter Stöcklein (Fraunhofer IBMT, Potsdam-Golm, Germany) for his excellent support regarding SPR measurements as well as Prof. Burkhard Micheel (University of Potsdam, Germany) for providing the Anti-FITC monoclonal antibody. Moreover, we thank Jörg Schenk (Hybrotec GmbH, UP Transfer GmbH, Potsdam-Golm, Germany) for delivering the DNA template encoding the Anti-FITC scFv as well as for the scientific support and many fruitful discussions. Plasmids encoding the membrane proteins Hb-EGF, EGFR, and ETB were kindly provided by Prof. Michael Schaefer (Universität Leipzig, Germany). We also wish to express our considerable thanks to Dr. Susanne Fenz (Universität Würzburg, Germany) and Prof. Thomas Schmidt (Leiden University, The Netherlands) for their collaboration regarding the GVU formation process. Our special thanks go to Dipl.-Translator Marie Burger, Dipl.-Chem. Devid Mrusek, and Dipl.-Biochem. Christian Hoffmeister (Fraunhofer IBMT, Potsdam-Golm, Germany) for their careful revision of this manuscript. This research is supported by the German Ministry of Education and Research (BMBF No. 0312039).

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Authors and Affiliations

  1. Fraunhofer IBMT, Branch Potsdam, Group of Cell-free Protein Synthesis, Am Mühlenberg 13, 14476, Potsdam, Germany

    Marlitt Stech, Andreas K. Brödel, Robert B. Quast, Rita Sachse & Stefan Kubick

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  1. Marlitt Stech
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Correspondence to Stefan Kubick .

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  1. Institut für Bioprozess- und Biosystemtechnik, Technische Universität Hamburg-Harburg, Hamburg, Germany

    An-Ping Zeng

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© 2013 Springer-Verlag Berlin Heidelberg

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Stech, M., Brödel, A.K., Quast, R.B., Sachse, R., Kubick, S. (2013). Cell-Free Systems: Functional Modules for Synthetic and Chemical Biology. In: Zeng, AP. (eds) Fundamentals and Application of New Bioproduction Systems. Advances in Biochemical Engineering/Biotechnology, vol 137. Springer, Berlin, Heidelberg. https://doi.org/10.1007/10_2013_185

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