Accession Number : ADA192123

Title :   New Approaches to Attenuated Hepatitis A Vaccine Development: Cloning and Sequencing of Cell-Culture Adapted Viral cDNA.

Descriptive Note : Annual rept. 15 Sep 85-14 Sep 86,

Corporate Author : NORTH CAROLINA UNIV AT CHAPEL HILL

Personal Author(s) : Lemon, Stanley M

PDF Url : ADA192123

Report Date : 13 Oct 1986

Pagination or Media Count : 25

Abstract : The goal of this research contract is to develop an improved understanding of the biological basis of cell culture adaptation and attenuation of Hepatitis A Vaccine (HAV), as this may ultimately lead to a rational approach to development of an effective attenuated vaccine. Accordingly, a cDNA genomic library spanning over 98% of the genome of a plaque-purified, cell culture-adapted variant of HM175 strain HAV (p17 ca-HM175) has been prepared in vector pBR322. Selected cDNA clones have been subcloned into the M13mp19 vector, and subjected to sequencing by the dideoxynucleotide method. In approximately 1 kb of the genome sequenced thus far, a total of 3 nucleotide substitutions have been identified in comparison with the known sequence of wild-type virus (wt-HM175); one of these mutations is silent, while the other two result in amino acid alterations in virus proteins. Completion of these studies will lead to an improved understanding of the biological basis of adaptation of virus to cell culture, and possibly attenuation. A secondary goal of the work in progress is to document the existence of strain-specific differences among various isolates of HAV, and to develop methods for the identification of specific HAV strains.

Descriptors :   *HEPATITIS, *VACCINES, ADAPTATION, AMINO ACIDS, CELLS(BIOLOGY), CLONES, CONTRACTS, CULTURES(BIOLOGY), GENETIC ENGINEERING, MUTATIONS, PROTEINS, VIRUSES, STRAINS(BIOLOGY), DEOXYRIBONUCLEIC ACIDS, HEPATITIS VIRUSES

Subject Categories : Pharmacology

Distribution Statement : APPROVED FOR PUBLIC RELEASE