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High Titre BacMam Viruses Improve Transduction Efficiency of Mammalian Cells
EP22426
Poster Title: High Titre BacMam Viruses Improve Transduction Efficiency of Mammalian Cells
Submitted on 16 Oct 2014
Author(s): Robert Possee, Elisabetta Locanto, Adam Chambers, Linda King
Affiliations: Oxford Expression Technologies Ltd, Oxford Brookes University
This poster was presented at MipTec
Poster Views: 2,092
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Poster Information
Abstract: Transient production of proteins in mammalian cells is fundamental to studies on gene function in health and disease. Many viral and plasmid vectors are available to enable the transfer of genes into mammalian cells, including baculovirus vectors.

Baculoviruses are insect-specific viruses that can transduce but not replicate in many mammalian cells. These BacMAM vectors utilise mammalian promoters to drive expression of target genes. One disadvantage of the current BacMAM system is that relatively high multiplicities of infection (50-200+ virus particles per cell) are often required for effective transduction.
This requires either concentration of the BacMAM virus (time-consuming/ labour intensive) or the use of chemical enhancers.

This study demonstrates that incorporation of a natural mutation in the FP25 gene into the BacMAM backbone vector can increase budded virus titres by several fold. Thus transduction efficacies can be improved without the need for concentration of virus or the use of chemical enhancers.
Summary: We present a method for producing high titre BacMam virus vectors for the efficient transduction of mammalian cells.References: Possee, RD, Hitchman, RB, Richards, KS, Mann, SG, Siaterli, E, Nixon, CP, Irving, CH, Assenberg, R, Alderton, D, Owens, RJ & King, LA (2008) Generation of baculovirus vectors for the high throughput production of proteins in insect cells. Biotechnol. Bioeng. 101 (6) 1115-22
Hitchman, RB, Possee, RD & King, LA (2012). High through put baculovirus expression in insect cells. Methods Mol Biol 824: 609-27
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