Human cytomegalovirus containing a .beta.-glucuronidase marker g

Chemistry: molecular biology and microbiology – Virus or bacteriophage – except for viral vector or...

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4353201, C12N 701, C12N 1538

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active

058770046

ABSTRACT:
This invention relates to a method for identifying non-essential genes of the human cytomegalovirus (HCMV) genome through the insertion of a .beta.-glucuronidase marker gene into a specified HCMV gene, such that, if the product of the HCMV gene is not expressed, the gene is identified as non-essential for replication of HCMV. This invention also relates to a method of screening for compounds which inhibit HCMV by the insertion of the .beta.-glucuronidase marker gene into a HCMV gene, such that the enzyme marker is expressed and cleaves a conjugate chemical substrate in an assay system to yield a detectable fluorescing product or to result in a color change. This invention further provides the gene responsible in HCMV early cytopathic effect.

REFERENCES:
patent: 5432081 (1995-07-01), Jefferson
Weston et al., "Sequence of the Short Unique Region, Short Repeats, and Part of the Long Repeats of Human Cytomegalovirus," J. Mol. Biol. , 192(2):pp. 177-208 (1986).
Gretch et al., "A Multigene Family Encodes the Human Cytomegalovirus Glycoprotein Complex gcII (gp47-52 Complex)," J. Virology, 62(6):pp. 1956-1962 (1988).
Spaete et al., "Insertion and deletion mutagenesis of the human cytomegalovirus genome," Proc. Natl. Acad. Sci. USA, 84(20):pp. 7213-7217 (1987).
Jefferson et al., ".beta.-Glucuronidase from Escherichia coli as a gene-fusion marker," Proc. Natl. Acad. Sci. USA, 83(22):pp. 8447-8451 (1986).
Jones et al., "A Cluster of Dispensable Genes within the Human Cytomegalovirus Genome Short Component: IRS1, US1-US5, and the US6 Family," J. Virology 66(4):pp. 2541-2546 (1992).
Jones et al., "Replacement Mutagenesis of the Human Cytomegalovirus Genome: US10 and US11 Gene Products are Non-essential," J. Virology 65(11):pp. 5860-5872 (1991).
Stasiak et al., "Transactivation of the Cytomegalovirus ICP 36 Gene Promotor Requires the .alpha. Gene Product TRS1 in Addition to IE1 and IE2," J. Virology, 66(2):pp. 1050-1058 (1992).

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