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August 17-18, 2017 | Toronto, Canada
ANNUAL BIOTECHNOLOGY CONGRESS
allied
academies
Ann Biol Sci, 2017
ISSN: 2348-1927
Poterszman Arnaud
Starsbourg University, France
Bottom-up strategies for reconstitution of multi-protein complexes using the
baculovirus expression system
T
he production of a homogeneous protein sample in
sufficient quantities is an essential prerequisite not only
for structural investigations but represents also a rate-limiting
step for many functional studies. In the cell, a large fraction of
eukaryotic proteins exists as large multicomponent assemblies
with many subunits, which act in concert to catalyze specific
activities. Genome editing allows to isolate native protein
complexes produced from their natural genomic contexts
but their limited natural abundance is often limited and so
recombinant expression and reconstitution are then required.
The
baculovirus
expression vector system (BEVS) has turned
out to be particularly powerful, unlocking the structure and
mechanism of many important complex assemblies that had
remained inaccessible to detailed analysis beforehand. Here,
we will comment on current developments and their potential
to accelerate protein complex research: Use of Lambda red
recombination in
E. coli
for manipulation and improvement
of the baculoviral genome, vector development for parallel
expression/co-expression screening and assembly of multi-
gene constructs from synthetic biology approaches. As model
systems, we will use human multi-protein complexes invoved
in the regulation of gene expression such the pTefb cdk/cyclin
pair, nuclear hormone receptor complexes or the 10 subunits
transcription/DNA repair complex TFIIH. We will describe state-
of-art strategies for the efficient production of multiprotein
complexes using the baculovirus/insect cell expression system.
Here, we will comment on current developments and their
potential toaccelerateproteincomplex research:Useof Lambda
red recombination in
E. coli
for manipulation and improvement
of the baculoviral genome, vector development for parallel
expression/co-expression screening and assembly of multi-
gene constructs from synthetic biology approaches. As model
systems, we will use human multi-subunit transcription factors
such as Cdk/cyclin pairs, nuclear hormone receptor complexes
or the 10 subunits transcription/DNA repair complex TFIIH.
Speaker Biography
Poterszman Arnaud, after studying at ENS Cachan, completed his PhD from Strasbourg
University and joined the CNRS one year later. He holds a CNRS Research Director po-
sition and performs his studies at the Department of Integrated Structural Biology at
IGBMC, Illkirch France. He has a dual expertise in Structural and Molecular Biology,
with insights on expression technologies and sample preparation. His research is fo-
cused on eukaryotic multi-protein complexes involved in transcription regulation and
DNA repair by nucleotide excision, particularly, the transcription/DNA repair factor TFI-
IH and its partners. He has around 50 publications in Pubmed, h-index 21.
e:
Arnaud.Poterszman@igbmc.frPoterszman Arnaud, Ann Biol Sci, 2017, 5:3
DOI: 10.21767/2348-1927-C1-001