Polydnavirus genes that enhance the baculovirus expression vector system.
The baculovirus expression vector system (BEVS) is a powerful and versatile system for protein expression, which has many advantages. However, a limitation of any lytic viral expression system, including BEVS, is that death and lysis of infected insect cells terminates protein production. This results in interruption of protein production and higher production costs due to the need to set up new infections, maintain uninfected cells, and produce pure viral stocks. Genetic methods to slow or prevent cell death while maintaining high-level, virus-driven protein production could dramatically increase protein yields. Several approaches have been used to improve the BEVS and increase the synthesis of functional proteins. Successful enhancement of the BEVS was obtained when various gene elements were added to the virus, secretion and posttranslational processing were modified, or protein integrity was improved. A gene family from the insect virus Campoletis sonorensis ichnovirus (CsIV) was discovered that delays lysis of baculovirus-infected cells, thereby significantly enhancing recombinant protein production in the BEVS system. By using the CsIV vankyrin gene family, protein production in the vankyrin-enhanced BEVS (VE-BEVS) was increased by a factor of 4- to 15-fold by either coexpressing the vankyrin protein from a dual BEVS or by providing its activity in trans by expressing the vankyrin protein from a stably transformed cell line. In sum, VE-BEVS is an enhancement of the existing BEVS technology that markedly improves protein expression levels while reducing the cost of labor and materials.
Fath-Goodin
Angelika
A
Department of Entomology, S-225 Agricultural Science Building North University of Kentucky, Lexington, Kentucky 40546, USA.
Kroemer
Jeremy
J
Martin
Stacy
S
Reeves
Krista
K
Webb
Bruce A
BA
D000595:Amino Acid Sequence; D000818:Animals; D017487:Ankyrins / Q000235:genetics; D016367:Baculoviridae / Q000235:genetics*; D002461:Cell Line, Transformed; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors / Q000235:genetics*; D007313:Insecta; D008969:Molecular Sequence Data; D018071:Polydnaviridae / Q000235:genetics*
D017487:Ankyrins