Development of a chitinase and v-cathepsin negative bacmid for improved integrity of secreted recombinant proteins

The application of the baculovirus-insect cell expression system for the production of integral membrane and secreted proteins is often more troublesome than for cytoplasmic proteins. One protein expressed at low levels in insect cells is the Theileria parva sporozoite surface protein p67. Theileria...

Full description

Saved in:
Bibliographic Details
Published in:Journal of virological methods 2004-12, Vol.122 (1), p.113-118
Main Authors: Kaba, Stephen A., Salcedo, Adriana M., Wafula, Paul O., Vlak, Just M., van Oers, Monique M.
Format: Article
Language:English
Subjects:
Baculoviridae - enzymology
Baculoviridae - genetics
Baculovirus
Baculovirus expression system
baculovirus system
Biological and medical sciences
Cathepsins - genetics
cattle
Chitinase
Chitinases - genetics
East Coast fever
escherichia-coli
expression
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation, Viral
Genetic Vectors
Honeybee mellitin signal
immunity
immunogenicity
insect cells
Melitten - genetics
Melitten - physiology
Microbiology
p67
Protein Sorting Signals
Protozoan Proteins - genetics
Protozoan Proteins - metabolism
Recombinant Proteins - metabolism
Techniques used in virology
Theileria parva
v-cathepsin
vaccine antigen
Virology
virus
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The application of the baculovirus-insect cell expression system for the production of integral membrane and secreted proteins is often more troublesome than for cytoplasmic proteins. One protein expressed at low levels in insect cells is the Theileria parva sporozoite surface protein p67. Theileria parva is a protozoan parasite, which causes the tick-transmitted disease East Coast fever in cattle. Baculovirus vectors were engineered to produce a secreted form of p67 by replacing the signal peptide of p67 with the honeybee mellitin signal sequence and deleting a putative membrane anchor from the C-terminus. Furthermore, the chitinase and v-cathepsin genes were deleted from the baculovirus expression vector in a bacmid setup, allowing broad scale application of this novel vector. Deletion of the chitinase and v-cathepsin gene had a positive effect on the integrity of both the intracellular and secreted recombinant protein.
ISSN:0166-0934
1879-0984
DOI:10.1016/j.jviromet.2004.07.006