Differentiation of Two Bovine Lentiviruses by a Monoclonal Antibody on the Basis of Epitope Specificity

Abstract

Bovine immunodeficiency virus (BIV) and

Unique Epitope of Bovine Immunodeficiency Virus Gag Protein Spans the Cleavage Site between p16 MA and p2L

“…The pATH gag-3 clone contains a 0.8-kb capsid gene from strain R29 of BIV (1) including MAb 10H1 (25). The protein of the pATH gag-3 clone expressed in E. coli starts with Gag residue R114, while the capsid protein starts with residue R149 (20).…”

“…A pATH gag-3 clone containing 781 bp of the gag-coding region from BIV strain R29 was used to generate BIV MAbs, including MAb 10H1. The production and characterization of MAb 10H1 were described earlier (25). One 751-bp fragment of the gag gene insert was subcloned into prokaryotic expression vector pQE32, designated pQE32-gag (25,26), and was used as a template for PCR amplification of gag gene fragments in this study.…”

“…We designed primers with sequences specific within the 163-bp DNA region corresponding to the 6.4-kDa protein (25). For cloning of BIV gag fragments into the pQE32 expression vector (QIA-GEN), the forward primers had BamHI restriction sites at their 5Ј ends, and the reverse primers had HindIII sites at their 3Ј ends.…”

“…The BIV gag gene has been cloned, and monoclonal antibodies (MAbs) to recombinant Gag protein have been developed (22,25,26). One of our MAbs, designated 10H1, reacts specifically with the BIV Gag protein but not with the JDV Gag protein, indicating that BIV has at least one epitope different from JDV epitopes.…”

Immunological Detection of Avian Influenza Virus in Infected Ducks by Monoclonal Antibodies Against AIV-H5N1

“…Western blotting assay was carried out according to procedure described by Zheng et al (2001). Formaldehyde inactivated AIV-H5N1 were diluted in an equal volume of sample loading buffer (1.3% SDS, 5% mercaptoethanol, 0.0625 M Tris-HCl pH 6.8, 10% glycerol, 0.001% bromophenol blue).…”