2. INTRODUCTION
The baculovirus vector system is widely used for the expression of recombinant proteins in
cultured insect cells.
It is one of the most versatile and powerful systems for eukaryotic expression of
recombinant proteins.
This system is particularly advantageous for large-scale preparation of proteins that require
expression in eukaryotic host cells.
In these cases, prokaryotic expression systems are often inadequate and the baculovirus
expression system could be a good alternative.
The shuttle vector is called bacmid, and gene expression is under the control of a strong
promoter pPolh.
Baculovirus has also been used with mammalian cell lines known as BacMam system.
Baculovirus –
• Envelop, double –stranded DNA
virus
• Belongs to baculoviridae family
• Most widely used baculovirus
species is Autographa Californica
multiple nucleopolyhedrovirus
(AcMNPV)
• Mostly infects Lepidopterans
(moths and butterflies) such as
Spodoptera frugiperda and Bmbyx
mori. Image credit - Berretta, M. F. , Baculovirus gene expression.
4. BACMID
• Shuttle vector
• The bacmid is essentially a very large
plasmid containing the baculovirus
genome modified to carry a lacZ gene.
• attTn7 (a short segment containing the
attachment site for the bacterial
transposon Tn7 ) inserted in the lacZ
coding region.
• Kanamycin resistance marker
• A low copy number mini- fertily replicon.
• Transposition is the process by which
genetic elements move between different
locations of the genome.
https://en.vectorbuilder.com/resources/vector-
system/pBV.html
Image courtesy -
5. Image credits - Sui, Dong & Wang, Kuanquan & Park, Heemin & Chae, Jinseok. (2014). Bright field
microscopic cells counting method for BEVS using nonlinear convergence index sliding band filter.
Biomedical engineering online. 13. 147. 10.1186/1475-925X-13-147.
6. PROTOCOL
Serial Passage –
• Serial passage is the
process of growing
bacteria or a virus in
iterations or repetition.
• A virus may be grown in
one environment, and
then a portion of that virus
population can be
removed and put into a
new environment.
• P0 – The virus progeny
from a transfection or
cotransfection.
• P1- Second generation of
the virus
7. References
Mena, J. A. , Kamen, A. A. , Insect cell technology is a versatile
and robust vaccine manufacturing platform. Expert Rev. Vaccines
2011, 10, 1063–1081.
Hu, Y. C. , Baculovirus as a highly efficient expression vector in
insect and mammalian cells. Acta Pharmacol. Sin. 2005, 26,
405–416.
Wickham, T. J. , Davis, T. , Granados, R. R. , Shuler, M. L. ,
Wood, H. A. , Screening of insect cell lines for the production of
recombinant proteins and infectious virus in the baculovirus
expression.
https://www.news-medical.net/life-sciences/The-Baculovirus-
Expression-Vector-System-(BEVS).aspx
https://www.sciencedirect.com/topics/medicine-and-
dentistry/baculovirus-expression-system
https://en.vectorbuilder.com/resources/vector-system/pBV.html