Methods for identifying non-essential genes of the human cytomeg

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage

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435 6, C12Q 170, C12Q 168

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058436375

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: 5026635 (1991-06-01), Ferguson et al.
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):177-208 (1986).
Gretch et al., "A Multigene Family Encodes the Human Cytomegalovirus Glycoprotein Complex gcII (gp47-52 Complex)," J. Virology, 62(6):1956-1962 (1988).
Spaete et al., "Insertion and deletion mutagenesis of the human cytomegalovirus genome," Proc. Natl. Acad. Sci. USA, 84(20):7213-7217 (1987).
Jefferson et al., ".beta.-Glucuronidase from Escherichia coli as a gene-fusion marker," Proc. Natl. Acad. Sci. USA, 83(22):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):2541-46 (1992).
Jones et al., "Replacement Mutagenesis of the Human Cytomegalovirus Genome: US10 and US11 Gene Products are Non-essential," J. Virology 65(11):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):1050-1058 (1992).
Takekoshi et al. (1991) Gene 101:209-213, May 1991.
Jefferson et al. (1986) Proc. Natl. Acad. Sci. USA 83:8447-8451, Nov. 1986.

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