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[Preparation of P52 recombinant antigenic protein from human cytomegalovirus (HCMV)].

Abstract

Analysis of published reports helped us single out the most potent antigens among HCMV proteins: phosphoproteins pp150(UL32) and p52(UL44). Theoretical computer analysis of p52 epitopes showed the main antigenic determinants not cross-reacting with antigens of other viruses. Virus-containing (strain AD169) material was obtained and genome DNA was isolated. Amplification of a site of gene UL44 coding for unique determinants detected a PCR fragment of required electrophoretic mobility. The fragment was cloned in vector pLBE. The specificity of cloning was confirmed by restriction analysis of theoretical sites. Nucleotide sequence of cloned fragment of UL44 gene was studied by Maxam-Gilbert's method. Cloning in expressing bacterial vectors helped obtain HCMV recombinant protein p52 in the pure form and fused with beta-galactosidase. Enzyme immunoassay with HCMV-positive and negative donor sera and ABBOTT HCMV sera showed that recombinant p52 increased the sensitivity and specificity of a previously obtained recombinant pp150 as an antigen to HCMV-IgG and HCMV-IgM. The sensitivity and specificity is 100% with 98-99% reliability.

Authors+Show Affiliations

Department of Molceular Biology, Faculty of Natural Sciences, Novosibirsk State University.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

English Abstract
Journal Article

Language

rus

PubMed ID

11186455

Citation

Susloparov, M A., et al. "[Preparation of P52 Recombinant Antigenic Protein From Human Cytomegalovirus (HCMV)]." Molekuliarnaia Genetika, Mikrobiologiia I Virusologiia, 2000, pp. 24-9.
Susloparov MA, Susloparov IM, Pliasunov IV, et al. [Preparation of P52 recombinant antigenic protein from human cytomegalovirus (HCMV)]. Mol Gen Mikrobiol Virusol. 2000.
Susloparov, M. A., Susloparov, I. M., Pliasunov, I. V., Bakhtina, M. M., Safronov, P. F., Grishaev, M. P., Blinov, V. M., & Ivan'kina, T. I. u. (2000). [Preparation of P52 recombinant antigenic protein from human cytomegalovirus (HCMV)]. Molekuliarnaia Genetika, Mikrobiologiia I Virusologiia, (4), 24-9.
Susloparov MA, et al. [Preparation of P52 Recombinant Antigenic Protein From Human Cytomegalovirus (HCMV)]. Mol Gen Mikrobiol Virusol. 2000;(4)24-9. PubMed PMID: 11186455.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Preparation of P52 recombinant antigenic protein from human cytomegalovirus (HCMV)]. AU - Susloparov,M A, AU - Susloparov,I M, AU - Pliasunov,I V, AU - Bakhtina,M M, AU - Safronov,P F, AU - Grishaev,M P, AU - Blinov,V M, AU - Ivan'kina,T Iu, PY - 2001/2/24/pubmed PY - 2001/3/3/medline PY - 2001/2/24/entrez SP - 24 EP - 9 JF - Molekuliarnaia genetika, mikrobiologiia i virusologiia JO - Mol Gen Mikrobiol Virusol IS - 4 N2 - Analysis of published reports helped us single out the most potent antigens among HCMV proteins: phosphoproteins pp150(UL32) and p52(UL44). Theoretical computer analysis of p52 epitopes showed the main antigenic determinants not cross-reacting with antigens of other viruses. Virus-containing (strain AD169) material was obtained and genome DNA was isolated. Amplification of a site of gene UL44 coding for unique determinants detected a PCR fragment of required electrophoretic mobility. The fragment was cloned in vector pLBE. The specificity of cloning was confirmed by restriction analysis of theoretical sites. Nucleotide sequence of cloned fragment of UL44 gene was studied by Maxam-Gilbert's method. Cloning in expressing bacterial vectors helped obtain HCMV recombinant protein p52 in the pure form and fused with beta-galactosidase. Enzyme immunoassay with HCMV-positive and negative donor sera and ABBOTT HCMV sera showed that recombinant p52 increased the sensitivity and specificity of a previously obtained recombinant pp150 as an antigen to HCMV-IgG and HCMV-IgM. The sensitivity and specificity is 100% with 98-99% reliability. SN - 0208-0613 UR - https://www.unboundmedicine.com/medline/citation/11186455/[Preparation_of_P52_recombinant_antigenic_protein_from_human_cytomegalovirus__HCMV_]_ L2 - https://antibodies.cancer.gov/detail/CPTC-NFKB2-1 DB - PRIME DP - Unbound Medicine ER -