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IRES mediated expression of viral 3C protease for enhancing the yield of FMDV empty capsids using baculovirus system

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Title IRES mediated expression of viral 3C protease for enhancing the yield of FMDV empty capsids using baculovirus system
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Creator Vivek Srinivas VM, Basagoudanavar SH, Hosamani M*
 
Subject Baculovirus expression
FMD virus
 
Description Not Available
For expression of FMDV empty capsids, high protease activity associated with 3C co-expressed with P1 polyprotein has been reported to adversely affect the yields of capsids. Limiting the levels of 3Cpro relative to P1-2A polypeptide is thus critical to enhance the yields. In this study, FMDV internal ribosome entry site (IRES) sequence which serves as an alternative to the CAP-dependent translation initiation mechanism, was used for controlled translation of 3C protease. Baculovirus expressing bicistronic cDNA cassette containing two open reading frames-FMDV capsid gene (P1-2A) and 3Cpro intervened by IRES was prepared. Analysis of the expression in insect cells infected with baculovirus showed increased accumulation of processed capsids. Recombinant capsids showed higher immunoreactivity similar to the whole virus antigen, when reacted with polyclonal antibodies against the purified whole virus 146S particles. Thus, inclusion of the IRES upstream of 3Cpro facilitated reduced expression of the protease in baculovirus expression system, without causing significant proteolysis, thereby contributing to improved yields of the processed capsid antigens.
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Date 2023-05-29T10:47:03Z
2023-05-29T10:47:03Z
2016-03-01
 
Type Article
 
Identifier Vivek Srinivas VM, Basagoudanavar SH, Hosamani M. IRES mediated expression of viral 3C protease for enhancing the yield of FMDV empty capsids using baculovirus system. Biologicals. 2016 Mar;44(2):64-8.
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http://krishi.icar.gov.in/jspui/handle/123456789/77845
 
Language English
 
Relation Not Available;
 
Publisher Elsevier