Project Details
Abstract
Epstein-Barr virus (EBV) is a human herpesvirus that is often implicated in
lymphoid and epithelial malignancies. Although EBV contains genes that are critical to
enabling the virus to retain its latency following infection, most of the EBV genes are
associated with lytic development. Furthermore, EBV has a large genome, so studying EBV
genes can be difficult, explaining why the functions of many EBV genes remain
unknown. This laboratory use a bacterial transposon, EZ::Tn<KAN-2>, and generated
about 50000 EBV mutants. The mutant DNA can be manipulated in Escherichia coli and
then transfected into cells for genetic and functional analyses. This investigation will use
these mutants to identify the genes that are involved in EBV maturation. The proposed
work will screen the mutants for those that cannot complete the maturation process;
characterize the functions of the genes that are necessary for maturation, and confirm
them by genetic complementation and electron microscopy. Furthermore, this proposed
study will focus on one of the EBV maturation events – nuclear egress. For EBV, BFLF2
has a sequence homology to one of the genes involved in nuclear egress of herpes
simplex virus type 1 (HSV-1). Since proteins participate in HSV-1 nuclear egress often
interact, the BFLF2 may be used as bait to identify the proteins that are crucial to EBV
nuclear egress by yeast two-hybrid screening. The interaction can be verified by GSTpull
down and coimmunoprecipitation. This investigation will ultimately identify the
genes that are required for EBV maturation; a process that is critical to EBV lytic
development and understanding how the virus proliferates in human cells.
Project IDs
Project ID:PA9405-0117
External Project ID:NSC94-3112-B182-004
External Project ID:NSC94-3112-B182-004
Status | Finished |
---|---|
Effective start/end date | 01/05/05 → 30/04/06 |
Keywords
- Epstein-Barr virus
- mutants
- maturation
- nuclear egress
- lytic cycle
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