Expression of the glycoprotein H of murine cytomegalovirus and identification of an N-terminal antibody-binding region

J. Xu, Tony Scalzo, P.A. Lyons, Geoffrey Shellam

Research output: Contribution to journalArticle

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Abstract

A series of overlapping fragments spanning the entire coding sequence of the gH gene of murine cytomegalovirus (MCMV) were expressed in Escherichia coil as fusion proteins with glutathione S-transferase (GST) using the pGEX expression system. A region of antibody-binding was mapped to the NH2-terminus of glycoprotein H (gH) between amino acid residues 26 and 90 on the basis of the reactivity of GST-gH fusion proteins with polyclonal antibodies to MCMV in Western blot analysis. Antibodies to gH were generated in mice immunized with the GST-gH fusion protein SK and shown to react with an 87-kDa polypeptide present in virion particles which was conserved in MCMV isolates obtained from diverse locations. They also recognized the gH protein in MCMV-infected cells, as well as gH expressed in Chinese Hamster Ovary cells. The antibodies to gH had a significant ELISA titer but no neutralizing activity in vitro. The antibody response to the GST-gH fusion protein did not modify the level of MCMV replication in the spleens of mice. (C) 1994 Academic Press, Inc.
Original languageEnglish
Pages (from-to)466-470
JournalVirology
Volume204
DOIs
Publication statusPublished - 1994

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Muromegalovirus
Glycoproteins
Antibodies
Glutathione Transferase
Proteins
Escherichia
Cricetulus
Virion
Antibody Formation
Ovary
Spleen
Western Blotting
Enzyme-Linked Immunosorbent Assay
Amino Acids

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Xu, J. ; Scalzo, Tony ; Lyons, P.A. ; Shellam, Geoffrey. / Expression of the glycoprotein H of murine cytomegalovirus and identification of an N-terminal antibody-binding region. In: Virology. 1994 ; Vol. 204. pp. 466-470.
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Expression of the glycoprotein H of murine cytomegalovirus and identification of an N-terminal antibody-binding region. / Xu, J.; Scalzo, Tony; Lyons, P.A.; Shellam, Geoffrey.

In: Virology, Vol. 204, 1994, p. 466-470.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Expression of the glycoprotein H of murine cytomegalovirus and identification of an N-terminal antibody-binding region

AU - Xu, J.

AU - Scalzo, Tony

AU - Lyons, P.A.

AU - Shellam, Geoffrey

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N2 - A series of overlapping fragments spanning the entire coding sequence of the gH gene of murine cytomegalovirus (MCMV) were expressed in Escherichia coil as fusion proteins with glutathione S-transferase (GST) using the pGEX expression system. A region of antibody-binding was mapped to the NH2-terminus of glycoprotein H (gH) between amino acid residues 26 and 90 on the basis of the reactivity of GST-gH fusion proteins with polyclonal antibodies to MCMV in Western blot analysis. Antibodies to gH were generated in mice immunized with the GST-gH fusion protein SK and shown to react with an 87-kDa polypeptide present in virion particles which was conserved in MCMV isolates obtained from diverse locations. They also recognized the gH protein in MCMV-infected cells, as well as gH expressed in Chinese Hamster Ovary cells. The antibodies to gH had a significant ELISA titer but no neutralizing activity in vitro. The antibody response to the GST-gH fusion protein did not modify the level of MCMV replication in the spleens of mice. (C) 1994 Academic Press, Inc.

AB - A series of overlapping fragments spanning the entire coding sequence of the gH gene of murine cytomegalovirus (MCMV) were expressed in Escherichia coil as fusion proteins with glutathione S-transferase (GST) using the pGEX expression system. A region of antibody-binding was mapped to the NH2-terminus of glycoprotein H (gH) between amino acid residues 26 and 90 on the basis of the reactivity of GST-gH fusion proteins with polyclonal antibodies to MCMV in Western blot analysis. Antibodies to gH were generated in mice immunized with the GST-gH fusion protein SK and shown to react with an 87-kDa polypeptide present in virion particles which was conserved in MCMV isolates obtained from diverse locations. They also recognized the gH protein in MCMV-infected cells, as well as gH expressed in Chinese Hamster Ovary cells. The antibodies to gH had a significant ELISA titer but no neutralizing activity in vitro. The antibody response to the GST-gH fusion protein did not modify the level of MCMV replication in the spleens of mice. (C) 1994 Academic Press, Inc.

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