Poster presentation
186
COMPONENTS OF THE BAC-TO-BAC® BACULOVIRUS
EXPRESSION SYSTEM
M. Shukurjonov, Sh. Khasanov, J. Abdurakhmanov, O. Ashirov,
S. Gaynazarova, N. Tosheva, S. Sasmakov, Sh. Azimova
S.Yu. Yunusov Institute of the Chemistry of Plant Substances Academy of sciences of the
Republic of Uzbekistan st. Mirzo-Ulugbek, 77, 100170 Tashkent
e-mail: genlab_icps@yahoo.com
Nowadays, the Bac-to-Bac® Baculovirus Expression System facilitates rapid and
efficient generation of recombinant baculoviruses. Recombinant
proteins produced by
baculovirus expression systems are almost indistinguishable from their natural analogs
by their functional properties, i.e. they have the correct post-translational modification,
and at the same time this system is characterized by high quantitative product yield.
Using insect baculoviruses cells for protein expression is safe for humans or other
mammals, since they are not pathogenic [1-2].
The Bac-to Bac® Baculovirus
Expression System takes advantage of the site-specific transposition
properties of the
Tn7 transposon to simplify and enhance the process of generating recombinant bacmid
DNA.
In our experiments, we started selecting the DH10Bac™ Escherichia coli strain to be
used as a host for our System component pFastBac™ vector. DH10Bac™ cells contain
a baculovirus vector (bacmid) with a mini-attTn7 targeting site and a helper plasmid.
After the pFastBac™ expression plasmid is transformed into DH10Bac™ cells,
transposition occurs between the mini-Tn7 element in the pFastBac™ vector and the
mini-attTn7 target site in the bacmid to generate recombinant bacmid. This
transposition reaction takes place in the presence of transposition proteins provided by
the helper plasmid.
This greatly simplifies subsequent research.
References:
1. Kajikawa M, Sasaki-Tabata K, Fukuhara H, Horiuchi M, Okabe Y, et al.
Silkworm Baculovirus Expression System for Molecular Medicine. 2012,
J Biotechnol
Biomaterial
S9:005. http://dx.doi.org/10.4172/2155-952X.S9-005.
2. Wu X, Kamei K, Sato H, Sato SI, Takano R, et al. High level expression of human
acidic fibroblast growth factor and basic fibroblast growth factor in silkworm (Bombyx
mori) using recombinant aculovirus.
Protein Expr Purif.
2001, 21: 192-200.
http://www.ncbi.nlm.nih.gov/pubmed/11162406
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