Characterization of pseudorabies virus glycoprotein gII expressed by recombinant baculovirus

Xuan, Xuenan; Nakamura, Toshihiro; Ihara, Takeshi; Sato, Ichiro; Tuchiya, Kotaro; Nosetto, Edgardo Omar; Ishihama, Akira; Ueda, Seinosuke

doi:10.1016/0168-1702(94)00112-p

Cited by 26 publications

(12 citation statements)

References 21 publications

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“…The structure of recombinant plasmid pBP50t was checked by restriction enzyme analysis, and the nucleotide sequence was analyzed with a model ABI 3100 DNA sequencer (Applied Biosystems, Foster City, Calif.). Construction of the recombinant baculovirus AcP50t was performed as described previously (23)(24)(25).…”

Section: Methodsmentioning

confidence: 99%

High-Level Expression of Truncated Surface Antigen P50 of Babesia gibsoni in Insect Cells by Baculovirus and Evaluation of Its Immunogenicity and Antigenicity

Fukumoto

Kadota

et al. 2003

Clin Vaccine Immunol

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Section: Methodsmentioning

confidence: 99%

High-Level Expression of Truncated Surface Antigen P50 of Babesia gibsoni in Insect Cells by Baculovirus and Evaluation of Its Immunogenicity and Antigenicity

Fukumoto

Kadota

et al. 2003

Clin Vaccine Immunol

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“…The structure of recombinant plasmid pBP50 was checked by restriction enzyme analysis. Construction of a recombinant baculovirus carrying the P50 gene (AcP50) was performed as described previously (22,23,24).…”

Section: Methodsmentioning

confidence: 99%

Identification and Expression of a 50-Kilodalton Surface Antigen of Babesia gibsoni and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

et al. 2001

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“…Sf9 cells were infected at 10 PFU/cell with a recombinant baculovirus carrying the EMA-1 gene (AcEMA-1), constructed as described above, or with a control recombinant baculovirus carrying the lacZ gene (AcLacZ) (20). After incubation for 4 days, cell extracts and culture media were tested by Western blotting with anti-EMA-1 serum.…”

Section: Resultsmentioning

confidence: 99%

Expression of Babesia equi Merozoite Antigen 1 in Insect Cells by Recombinant Baculovirus and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

et al. 2001

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The gene encoding the entire Babesia equi merozoite antigen 1 (EMA-1) was inserted into a baculovirus transfer vector, and a recombinant virus expressing EMA-1 was isolated. The expressed EMA-1 was transported to the surface of infected insect cells, as judged by an indirect fluorescent-antibody test (IFAT). The expressed EMA-1 was also secreted into the supernatant of a cell culture infected with recombinant baculovirus. Both intracellular and extracellular EMA-1 reacted with a specific antibody in Western blots. The expressed EMA-1 had an apparent molecular mass of 34 kDa that was identical to that of native EMA-1. The secreted EMA-1 was used as an antigen in an enzyme-linked immunosorbent assay (ELISA). The ELISA differentiated B. equi-infected horse sera from Babesia caballi-infected horse sera or normal horse sera. The ELISA was more sensitive than the complement fixation test and IFAT. These results demonstrated that the recombinant EMA-1 expressed in insect cells might be a useful diagnostic reagent for detection of antibodies to B. equi.Babesia equi is a tick-borne hemoprotozoan parasite that causes piroplasmosis in horses. Equine piroplasmosis is an economically important disease that is characterized by fever, anemia, and icterus and that is mostly prevalent in tropical and subtropical areas as well as in temperate climatic zones (15). Areas of endemicity include many parts of Europe, Africa, Arabia, and Asia (15). Due to the almost worldwide distribution of various tick vectors, the introduction of a carrier into areas of nonendemicity should be prevented.The complement fixation test (CFT) and indirect fluorescent-antibody test (IFAT) have commonly been used to detect B. equi infection. However, these serologic tests are generally restricted by the antibody detection limits and cross-reactivity (4, 5). Besides CFT and IFAT, the enzyme-linked immunosorbent assay (ELISA) with B. equi lysate antigen has been used for detection of antibodies to B. equi (19). However, the ELISA is hindered by a limited antigen supply and poor specificity (4,5,19).Merozoite antigen 1 (EMA-1) is the major surface protein of B. equi (8). It is considered an important candidate with which to develop a diagnostic reagent for detection of antibodies to B. equi (9, 10). A competitive inhibition ELISA (CI-ELISA) that can detect antibodies to B. equi based on a monoclonal antibody to EMA-1 has been developed by Knowles et al. (11), who demonstrated that it can be more sensitive than CFT in detecting antibodies to B. equi. The CI-ELISA offers the advantage of a high degree of specificity but the disadvantage of the requirement of a complicated operating procedure. Therefore, there is a need to develop a simple ELISA method.Here, we established a highly specific, sensitive, and simple ELISA method using recombinant EMA-1 expressed in insect cells by baculovirus. Our data indicated that the recombinant baculovirus-expressed EMA-1 should be a useful diagnostic reagent for detection of antibodies to B. equi in horses. MATERIALS AND MET...

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Characterization of pseudorabies virus glycoprotein gII expressed by recombinant baculovirus

Cited by 26 publications

References 21 publications

High-Level Expression of Truncated Surface Antigen P50 of Babesia gibsoni in Insect Cells by Baculovirus and Evaluation of Its Immunogenicity and Antigenicity

High-Level Expression of Truncated Surface Antigen P50 of Babesia gibsoni in Insect Cells by Baculovirus and Evaluation of Its Immunogenicity and Antigenicity

Identification and Expression of a 50-Kilodalton Surface Antigen of Babesia gibsoni and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

Expression of Babesia equi Merozoite Antigen 1 in Insect Cells by Recombinant Baculovirus and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

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