PubMed:23619971
Annnotations
{"target":"http://pubannotation.org/docs/sourcedb/PubMed/sourceid/23619971","sourcedb":"PubMed","sourceid":"23619971","source_url":"https://www.ncbi.nlm.nih.gov/pubmed/23619971","text":"High-yield production of the VP1 structural protein epitope from serotype O foot-and-mouth disease virus in Escherichia coli.\nFor effective control of foot-and-mouth disease (FMD), the development of rapid diagnostic systems and vaccines are required against its etiological agent, FMD virus (FMDV). To accomplish this, efficient large-scale expression of the FMDV VP1 protein, with high solubility, needs to be optimized. We attempted to produce high levels of a serotype O FMDV VP1 epitope in Escherichia coli. We identified the subtype-independent serotype O FMDV VP1 epitope sequence and used it to construct a glutathione S-transferase (GST) fusion protein. For efficient production of the FMDV VP1 epitope fused to GST (VP1e-GST), four E. coli strains and three temperatures were examined. The conditions yielding the greatest level of VP1e-GST with highest solubility were achieved with E. coli BL21(DE3) at 25 °C. For high-level production, fed-batch cultures were conducted in 5-l bioreactors. When cells were induced at a high density and complex feeding solutions were supplied, approximately 11 g of VP1e-GST was obtained from a 2.9-l culture. Following purification, the VP1 epitope was used to immunize rabbits, and we confirmed that it induced an immune response.","tracks":[{"project":"Allie","denotations":[{"id":"SS1_23619971_1_0","span":{"begin":151,"end":173},"obj":"expanded"},{"id":"SS2_23619971_1_0","span":{"begin":175,"end":178},"obj":"abbr"},{"id":"SS1_23619971_1_1","span":{"begin":282,"end":291},"obj":"expanded"},{"id":"SS2_23619971_1_1","span":{"begin":293,"end":297},"obj":"abbr"},{"id":"SS1_23619971_4_0","span":{"begin":615,"end":640},"obj":"expanded"},{"id":"SS2_23619971_4_0","span":{"begin":642,"end":645},"obj":"abbr"},{"id":"SS1_23619971_5_0","span":{"begin":700,"end":724},"obj":"expanded"},{"id":"SS2_23619971_5_0","span":{"begin":726,"end":734},"obj":"abbr"}],"relations":[{"id":"AE1_23619971_1_0","pred":"abbreviatedTo","subj":"SS1_23619971_1_0","obj":"SS2_23619971_1_0"},{"id":"AE1_23619971_1_1","pred":"abbreviatedTo","subj":"SS1_23619971_1_1","obj":"SS2_23619971_1_1"},{"id":"AE1_23619971_4_0","pred":"abbreviatedTo","subj":"SS1_23619971_4_0","obj":"SS2_23619971_4_0"},{"id":"AE1_23619971_5_0","pred":"abbreviatedTo","subj":"SS1_23619971_5_0","obj":"SS2_23619971_5_0"}],"attributes":[{"subj":"SS1_23619971_1_0","pred":"source","obj":"Allie"},{"subj":"SS2_23619971_1_0","pred":"source","obj":"Allie"},{"subj":"SS1_23619971_1_1","pred":"source","obj":"Allie"},{"subj":"SS2_23619971_1_1","pred":"source","obj":"Allie"},{"subj":"SS1_23619971_4_0","pred":"source","obj":"Allie"},{"subj":"SS2_23619971_4_0","pred":"source","obj":"Allie"},{"subj":"SS1_23619971_5_0","pred":"source","obj":"Allie"},{"subj":"SS2_23619971_5_0","pred":"source","obj":"Allie"}]}],"config":{"attribute types":[{"pred":"source","value type":"selection","values":[{"id":"Allie","color":"#93ecce","default":true}]}]}}