Deletion of Marek's disease virus large subunit of ribonucleotide reductase impairs virus growth in vitro and in vivo. 2013

Aijun Sun, and Lucy F Lee, and Owais A Khan, and Mohammad Heidari, and Huanmin Zhang, and Blanca Lupiani, and Sanjay M Reddy
Department of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M University, College Station, TX 77843, USA.

Marek's disease virus (MDV), a highly cell-associated lymphotropic alphaherpesvirus, is the causative agent of a neoplastic disease in domestic chickens called Marek's disease (MD). In the unique long (UL) region of the MDV genome, open reading frames UL39 and UL40 encode the large and small subunits of the ribonucleotide reductase (RR) enzyme, named RR1 and RR2, respectively. MDV RR is distinguishable from that present in chicken and duck cells by monoclonal antibody T81. Using recombinant DNA technology we have generated a mutant MDV (Md5deltaRR1) in which RR1 was deleted. PCR amplification of the RR gene in Md5deltaRR1-infected duck embryo fibroblasts (DEF) confirmed the deletion of the 2.4 kb RR1 gene with a resultant amplicon of a 640-bp fragment. Restriction enzyme digests with SalI confirmed a UL39 deletion and the absence of gross rearrangement. The biologic characteristics of Md5deltaRR1 virus were studied in vitro and in vivo. The Md5deltaRR1 replicated in DEF, but significantly slower than parental Md5-BAC, suggesting that RR is important but not essential for replication in fibroblasts. In vivo studies, however, showed that the RR1 deletion virus was impaired for its ability to replicate in chickens. Inoculation of specific-pathogen-free (SPF) chickens with Md5deltaRR1 showed the mutant virus is nonpathogenic and does not induce MD in birds. A revertant virus, Md5deltaRR1/R, was generated with the restored phenotype of the parental Md5-BAC in vivo, indicating that RR is essential for replication of the virus in chickens. Protection studies in SPF chickens indicated that the Md5deltaRR1 virus is not a candidate vaccine against MD.

UI MeSH Term Description Entries
D002478 Cells, Cultured Cells propagated in vitro in special media conducive to their growth. Cultured cells are used to study developmental, morphologic, metabolic, physiologic, and genetic processes, among others. Cultured Cells,Cell, Cultured,Cultured Cell
D002642 Chick Embryo The developmental entity of a fertilized chicken egg (ZYGOTE). The developmental process begins about 24 h before the egg is laid at the BLASTODISC, a small whitish spot on the surface of the EGG YOLK. After 21 days of incubation, the embryo is fully developed before hatching. Embryo, Chick,Chick Embryos,Embryos, Chick
D002645 Chickens Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA. Gallus gallus,Gallus domesticus,Gallus gallus domesticus,Chicken
D004372 Ducks A water bird in the order Anseriformes (subfamily Anatinae (true ducks)) with a broad blunt bill, short legs, webbed feet, and a waddling gait. Duck
D000595 Amino Acid Sequence The order of amino acids as they occur in a polypeptide chain. This is referred to as the primary structure of proteins. It is of fundamental importance in determining PROTEIN CONFORMATION. Protein Structure, Primary,Amino Acid Sequences,Sequence, Amino Acid,Sequences, Amino Acid,Primary Protein Structure,Primary Protein Structures,Protein Structures, Primary,Structure, Primary Protein,Structures, Primary Protein
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012264 Ribonucleotide Reductases Ribonucleotide Reductase,Reductase, Ribonucleotide,Reductases, Ribonucleotide
D013047 Specific Pathogen-Free Organisms Animals or humans raised in the absence of a particular disease-causing virus or other microorganism. Less frequently plants are cultivated pathogen-free. Pathogen-Free Organisms,Specific Pathogen Free,Organism, Pathogen-Free,Organism, Specific Pathogen-Free,Organisms, Pathogen-Free,Organisms, Specific Pathogen-Free,Pathogen Free Organisms,Pathogen Free, Specific,Pathogen Frees, Specific,Pathogen-Free Organism,Pathogen-Free Organism, Specific,Pathogen-Free Organisms, Specific,Specific Pathogen Free Organisms,Specific Pathogen-Free Organism
D016133 Polymerase Chain Reaction In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short oligonucleotide flanking sequences (primers). The essential steps include thermal denaturation of the double-stranded target molecules, annealing of the primers to their complementary sequences, and extension of the annealed primers by enzymatic synthesis with DNA polymerase. The reaction is efficient, specific, and extremely sensitive. Uses for the reaction include disease diagnosis, detection of difficult-to-isolate pathogens, mutation analysis, genetic testing, DNA sequencing, and analyzing evolutionary relationships. Anchored PCR,Inverse PCR,Nested PCR,PCR,Anchored Polymerase Chain Reaction,Inverse Polymerase Chain Reaction,Nested Polymerase Chain Reaction,PCR, Anchored,PCR, Inverse,PCR, Nested,Polymerase Chain Reactions,Reaction, Polymerase Chain,Reactions, Polymerase Chain
D017384 Sequence Deletion Deletion of sequences of nucleic acids from the genetic material of an individual. Deletion Mutation,Deletion Mutations,Deletion, Sequence,Deletions, Sequence,Mutation, Deletion,Mutations, Deletion,Sequence Deletions

Related Publications

Aijun Sun, and Lucy F Lee, and Owais A Khan, and Mohammad Heidari, and Huanmin Zhang, and Blanca Lupiani, and Sanjay M Reddy
May 1994, Journal of virology,
Aijun Sun, and Lucy F Lee, and Owais A Khan, and Mohammad Heidari, and Huanmin Zhang, and Blanca Lupiani, and Sanjay M Reddy
September 2014, Archives of virology,
Aijun Sun, and Lucy F Lee, and Owais A Khan, and Mohammad Heidari, and Huanmin Zhang, and Blanca Lupiani, and Sanjay M Reddy
August 1993, The Journal of biological chemistry,
Aijun Sun, and Lucy F Lee, and Owais A Khan, and Mohammad Heidari, and Huanmin Zhang, and Blanca Lupiani, and Sanjay M Reddy
April 2007, Anti-cancer drugs,
Aijun Sun, and Lucy F Lee, and Owais A Khan, and Mohammad Heidari, and Huanmin Zhang, and Blanca Lupiani, and Sanjay M Reddy
December 1994, The Journal of general virology,
Aijun Sun, and Lucy F Lee, and Owais A Khan, and Mohammad Heidari, and Huanmin Zhang, and Blanca Lupiani, and Sanjay M Reddy
February 1990, Virology,
Aijun Sun, and Lucy F Lee, and Owais A Khan, and Mohammad Heidari, and Huanmin Zhang, and Blanca Lupiani, and Sanjay M Reddy
April 2007, Clinical cancer research : an official journal of the American Association for Cancer Research,
Aijun Sun, and Lucy F Lee, and Owais A Khan, and Mohammad Heidari, and Huanmin Zhang, and Blanca Lupiani, and Sanjay M Reddy
June 1992, The Journal of general virology,
Aijun Sun, and Lucy F Lee, and Owais A Khan, and Mohammad Heidari, and Huanmin Zhang, and Blanca Lupiani, and Sanjay M Reddy
January 1972, American journal of veterinary research,
Aijun Sun, and Lucy F Lee, and Owais A Khan, and Mohammad Heidari, and Huanmin Zhang, and Blanca Lupiani, and Sanjay M Reddy
January 2023, International journal of molecular sciences,
Copied contents to your clipboard!