A rapid profiling assay for avian leukosis virus subgroup E proviruses in chickens.
Endogenous retroviral elements (ERVs) are prolific components of the genomes of complex species, typically occupying more sequence space than do essential, protein-encoding genes. Much of what we know today about the structure and function, as well as the evolution and pathogenic potential, of ERVs was fleshed out over several decades during the last century using the avian leukosis virus subgroup E-related (ALVE) family of endogenous retroviruses of chickens as a model system. A critical enabling factor in the elucidation of ALVE structure and function is the ability to detect and unambiguously identify specific ALVE proviral elements and to develop accurate element profiles for individual chickens under study. Currently, the most common approach for ALVE locus detection involves element-specific PCR assays carried out using primers that target host DNA near the insertion site of the provirus (i.e., the upstream and downstream flanks of the unoccupied site). Here we describe a new approach for proviral detection that exploits restriction enzyme sites in flanking DNA to develop ALVE element profiles more rapidly than with assays currently in use. Moreover, unlike element-specific PCR tests, the "profiling" assay detects novel ALVEs for which insertion sites have not yet been identified as well as previously characterized elements.
Katherine
K
Rutherford
Nancy
N
McLean
Bernhard F
BF
Benkel
D000818:Animals; D001353:Avian Leukosis / Q000821:virology*; D001354:Avian Leukosis Virus / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D002645:Chickens; D004262:DNA Restriction Enzymes / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D011201:Poultry Diseases / Q000821:virology*; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D015183:Restriction Mapping / Q000379:methods* / Q000662:veterinary; D017422:Sequence Analysis, DNA / Q000662:veterinary
D004279:DNA, Viral; D004262:DNA Restriction Enzymes