Unknown

Dataset Information

0

Baculovirus expression of HCoV-OC43 nucleocapsid protein and development of a Western blot assay for detection of human antibodies against HCoV-OC43.


ABSTRACT: The nucleocapsid (N) gene of human coronavirus strain OC43 (HCoV-OC43) was amplified by reverse transcriptase-polymerase chain reaction, and cloned in pENTR/D-TOPO plasmid. This plasmid containing the N gene was recombined with in a BaculoDirect baculovirus DNA designed in order to express N protein in fusion with a C-terminal polyhistidine tag containing V5 epitope. Sf21 cells were transfected with recombinant baculovirus DNA. Recombinant N protein was extracted from infected cells, analysed by SDS-PAGE and Western blot, and purified by Ni2+ affinity procedure. Sera from 100 healthcare workers and five 2-3-year-old children were tested in a Western blot assay using the purified recombinant N protein. All of the sera from adults and two of the sera from children have a positive result.

SUBMITTER: Mourez T 

PROVIDER: S-EPMC7112889 | biostudies-literature | 2007 Feb

REPOSITORIES: biostudies-literature

altmetric image

Publications

Baculovirus expression of HCoV-OC43 nucleocapsid protein and development of a Western blot assay for detection of human antibodies against HCoV-OC43.

Mourez Thomas T   Vabret Astrid A   Han Yang Y   Dina Julia J   Legrand Loïc L   Corbet Sandrine S   Freymuth François F  

Journal of virological methods 20061031 2


The nucleocapsid (N) gene of human coronavirus strain OC43 (HCoV-OC43) was amplified by reverse transcriptase-polymerase chain reaction, and cloned in pENTR/D-TOPO plasmid. This plasmid containing the N gene was recombined with in a BaculoDirect baculovirus DNA designed in order to express N protein in fusion with a C-terminal polyhistidine tag containing V5 epitope. Sf21 cells were transfected with recombinant baculovirus DNA. Recombinant N protein was extracted from infected cells, analysed by  ...[more]

Similar Datasets

| S-EPMC371214 | biostudies-literature
| S-EPMC7901274 | biostudies-literature
| S-EPMC7112803 | biostudies-literature
| S-EPMC2168377 | biostudies-literature
| S-EPMC95681 | biostudies-literature
| S-EPMC8500843 | biostudies-literature
| S-EPMC7112824 | biostudies-literature
| S-EPMC7916441 | biostudies-literature
| S-EPMC5571433 | biostudies-other
| S-EPMC8344746 | biostudies-literature