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Title: CHARACTERIZATION OF AN OPEN READING FRAME IN THE GENOME OF MAREK'S DISEASE VIRUS COMMON TO ALL THREE SEROTYPES

Author
item Lee, Lucy
item Silva, Robert
item CUI, XIAOPING - MICHIGAN STATE UNIVERSITY
item REDDY, SANJAY - TEXAS A & M

Submitted to: International Marek's Disease Symposium Abstracts and Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 7/14/2004
Publication Date: 7/14/2004
Citation: Lee, L.F., Silva, R.F., Cui, X., Reddy, S. 2004. Characterization of an open reading frame (LORF11) in the genome of Marek's disease virus common to all three serotypes [abstract]. International Marek's Disease Symposium Abstracts and Proceedings. Paper No. I. 5.

Interpretive Summary:

Technical Abstract: DNA sequence data of the GA and Md5 strains of Marek's disease virus (MDV) revealed a large open reading frame (LORF11) located in the unique long region of MDV genome and comprising 2711 nucleotides in length and potentially encoding a protein of 903 amino acids. Sequence comparison between MDV serotypes revealed all three serotypes contained LORF11 homologs. No sequence homology was found with any other herpesviruses and therefore a unique MDV gene. The MDV-specific LORF11 gene was deleted from cosmid A6 by using the RecA-assisted restriction endonuclease cleavage method. The recombinant cosmid, A6(DLORF11), was transfected into DEF cells in conjunction with parental SN5, P89, SN16, and B40 cosmid clones. rMd5(DLORF11) plaques were evident 12'13 days after transfection. PCR amplification of DEF cells infected with mutant viruses confirmed the deletion of a 2.57-Kb fragment and resulted in a fragment of 296-bp fragment . Cosmid DNA RE digests with BamHI, EcoRI and HindIII confirmed the absence of rearrangement. The role of the deleted LORF 11 virus was studied in vitro and in vivo. In vitro growth characteristics of the rMd5/DLORF11 viruses were similar to those of parental rMd5, showing that LORF11 is not essential for replication in cultured cells. Preliminary in vivo studies showed that LORF11 is essential for replication in lymphoid organs. The significance of this result will be discussed in terms of tumor development.