Marek's disease virus modified for the expression of NDV-F protein | BCH-LMO-SCBD-48972 | Living Modified Organism | Biosafety Clearing-House


Living Modified Organism (LMO)

Decisions on the LMO Risk Assessments  
published: 20 Jun 2009 last updated: 26 Jan 2015
Living Modified Organism identity
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Marek's disease virus modified for the expression of NDV-F protein
Cellmune N
Recombinant Marek's disease virus (a.k.a Gallid herpesvirus 2) modified to express NDV-F protein gene to generate a vaccine that helps aid in the protection against Newcastle and Marek's diseases in poultry.
The term “Recipient organism” refers to an organism (either already modified or non-modified) that was subjected to genetic modification, whereas “Parental organisms” refers to those that were involved in cross breeding or cell fusion.
Marek's disease virus serotype 1 strain 207
Characteristics of the modification process
  • Electroporation
Some of these genetic elements may be present as fragments or truncated forms. Please see notes below, where applicable.
  • BCH-GENE-SCBD-103640-1 Glycoprotein B promoter | Gallid alphaherpesvirus 2
  • BCH-GENE-SCBD-103642-2 Transcription termination factor | Macaca mulatta polyomavirus 1 (SV40, Simian vacuolating virus 40, simian virus 40, Rhesus macaque polyomavirus)
  • BCH-GENE-SCBD-105090-4 Fusion protein gene | Avian orthoavulavirus 1 (Newcastle disease virus, NDV)
    Protein coding sequence | Production of medical or pharmaceutical compounds (human or animal) (Vaccines)
An insertion plasmid called pKA4BPF was engineered by cloning an expression cassette composed of gB promoter, F protein gene and transcription termination factor into the commercially available plasmid vector pUC119 and used for the development of the recombinant virus.

The gB promoter was used with the aim of effectively expressing the NDV-F protein gene (see below) in the cells infected with MDV1. It is known that the homologous UL28 of HSV1 functions by incorporating the virus DNA into viral particles. However, it is not known how the protein encoded by UL28 functions in MDV1.

Homologous recombination was conducted by transferring the insertion vector plasmid pKA4BPF into a chicken embryo primary cell line infected with the MDV1 CVI988 C17 strain, the recipient organism virus, based on electroporation.

The gB (glycoprotein B) promoter region was cloned from the CVI988 C17 strain of the Gallid herpesvirus 2. It is a 0.5kb fragment amplified through PCR, with the EcoRI site added at each 5' end. The gB promoter sequence is configured mostly with the 3'-terminal of UL28 gene, containing 20% of its ORFs.

NDV-F gene derived from the avirulent Newcastle disease virus (NDV) D26 strain.

Transcription termination factor is a 0.25kb fragment derived from the commercially available expression plasmid pSVL. It contains the polyA addition signal and also the 3'-terminal sequence (77 bases) of large T antigen ORF of SV40 and the 3'-terminal sequence (61 bases) of VP1, the major virus capsid.
LMO characteristics
  • Vaccine
Detection method(s)
Additional Information
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Risk Assessment generated by a regulatory process Living modified organism(s) 1