Expression of the Cloned Hemagglutinin-Neuraminidase Gene of the Newcastle Disease Virus in Mouse Myeloma Cells

Vaccination with autologous cancer cells expressing a potent foreign antigen is promising for immunotherapy of tumors. A construct was obtained to transfect cancer cells with the hemagglutinin-neuraminidase (HN) gene of the Newcastle disease virus (NDV). Specific primers were designed, and the HN cD...

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Published inMolecular biology (New York) Vol. 35; no. 5; pp. 785 - 791
Main Authors Risinskaya, N V, Fegeding, K V, Vasilenko, O V, Sudarikov, A B
Format Journal Article
LanguageEnglish
Published New York Springer Nature B.V 01.09.2001
Subjects
Allantoic fluid
Birds
Cancer
Cancer vaccines
Cell agglutination
double prime HN gene
Eggs
Enzyme-linked immunosorbent assay
Exo- alpha -sialidase
Expression vectors
Ice
Immunotherapy
Myeloma
Newcastle disease
Newcastle disease virus
Poultry
Primers
Proteins
RNA
Tumors
Vaccination
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Summary:Vaccination with autologous cancer cells expressing a potent foreign antigen is promising for immunotherapy of tumors. A construct was obtained to transfect cancer cells with the hemagglutinin-neuraminidase (HN) gene of the Newcastle disease virus (NDV). Specific primers were designed, and the HN cDNA was amplified from RNA isolated from the allantoic fluid of NDV-infected embryonated chicken eggs. The amplified fragment was cloned in pCR2.1, sequenced, and recloned in expression vector pCDNA3.1/Zeo(+). The resulting construct was used to transfect mouse myeloma cells SP2/0. Production of HN was checked by ELISA and by a neuraminidase activity assay. Cell agglutination on ice was proposed as a test for surface HN.[PUBLICATION ABSTRACT]
Bibliography:ObjectType-Article-2
SourceType-Scholarly Journals-1
ObjectType-Feature-1
content type line 23
ISSN:0026-8933
1608-3245
DOI:10.1023/A:1012346925499