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PMID: 7474071 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Constitutive episomal expression of polypeptide IX (pIX) in a 293-based cell line complements the deficiency of pIX mutant adenovirus type 5.

Journal of virology ·Vol. 69 ·No. 11 ·1995-11-00 ·Pages 6627-33

Caravokyri C, Leppard KN

Abstract

The human adenovirus type 5 capsid is composed of a number of distinct polypeptides. It has been shown previously that one of these, polypeptide IX (pIX), is not absolutely required for the production of viable virus. However, viruses lacking this polypeptide have a significantly reduced packaging limit and, in the one case studied, also show a thermolabile virion phenotype. This report describes the use of eukaryotic episomal vectors based on the Epstein-Barr virus replicon to generate cells which stably express pIX. These cells provide pIX that is efficiently incorporated into virions that are genetically pIX-; such enhanced thermostability. These cells have also been used to isolate a genetically pIX- virus having a genome of length some 2.3 kbp in excess of the previously defined packaging limit for pIX- virus; the resulting virions have wild-type thermostability. These cells expand the theoretical capacity of adenovirus vectors for foreign DNA to around 9.2 kbp and may therefore be useful in gene therapy applications in which vector capacity is limiting.

MeSH Terms
Adenoviruses, Human/genetics,metabolism Base Sequence Capsid/biosynthesis Capsid Proteins Cell Line DNA Primers DNA, Viral/metabolism Gene Expression Genetic Vectors HeLa Cells Herpesvirus 4, Human/genetics Humans Kinetics Molecular Sequence Data Plasmids Polymerase Chain Reaction Promoter Regions, Genetic RNA, Messenger/analysis,biosynthesis Replicon Restriction Mapping Transfection
Chemicals
Capsid Proteins DNA Primers DNA, Viral RNA, Messenger hexon capsid protein, Adenovirus
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Caravokyri C
Department of Biological Sciences, Universiy of Warwick, Coventry, United Kingdom.
Leppard K N
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.69.11.6627-6633.1995
Published
1995-11-00
Pages
6627-33
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC189571
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.69.11.6627-6633.1995 PMC Full Text View journal details
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