Identification and localization of reiterated sequences in the Choristoneura fumiferana MNPV genome.

Abstract

The genome of Choristoneura fumiferana nuclear polyhedrosis virus (CfMNPV) contained reiterated sequences interdispersed in four locations. These regions, termed RS, were found in EcoRI fragments A, F, E and B. The sequences were identified by hybridization of the fragment EcoRI-A to a Southern blot of EcoRI-digested viral DNA. Further confirmation and more precise localization of the RS sequences was obtained by hybridization of nick-translated 2P-labeled EcoRI-E fragment to Southern blots of viral DNA digested with EcoRI, BamHI, XbaI and BglIl. Hybridization of 32P-labeled EcoRI-E to HindIII blots of viral DNA revealed the presence of a 'ladder' consisting of eight fragments. The three fragments of the ladder with the lowest sizes represented the HindIII fragments, 0, PQ and R. The other five fragments were submolar in amount, in that they could not be seen in ethidium bromide-stained gels and probably represented minor virus variants that arose after passage of virus in larvae. Each variant was distinguished from the others by an additional insertion of 210 bp into the EcoRl-B fragment of the genome.

A Physical Map of the Oryctes Baculovirus Genome

“…Smith & Summers (1982) Ours is the third report of reiterated sequences in a baculovirus genome. A. caliJbrnica NPV was reported to have five regions of reiteration (Cochran & Faulkner, 1983) and C. fumiferana four regions (Arif & Doerfler, 1984). As yet, no function has been found associated with these regions.…”

“…[rugiperda NPV (Loh et al, 1981), Mamestra brassicae NPV (Wiegers & Vlak, 1984), Heliothis zea NPV (Knell & Summers, 1984), Orygia pseudotsugata NPV (Leisy et al, 1984)and ChoristoneurafumiJerana NPV (Arif & Doerfler, 1984). To orientate these circular genomes with respect to one another, the polyhedrin genes, which contain conserved sequences, have been designated as the starting points of baculovirus maps (Vlak & Smith, 1982).…”

Molecular Cloning and Physical Mapping of the Genome of Bombyx Mori Nuclear Polyhedrosis Virus

“…As expected, several EcoRI fragments less than 400 bp were identified on a 1.5 ~o agarose gel (data not shown). These areas containing several smaller EcoRI fragments seemed to be the socalled repeated sequences also found in AcNPV (Erlandson et al, 1984;Guarino et al, 1986) and other baculoviruses (Kuzio & Faulkner, 1984;Arif & Doerfler, 1984). Three repeated sequences could not be mapped exactly; however, their locations are at either end or on both ends of the small EcoRI N, P, and R fragments.…”

A Baculovirus (Bombyx mori Nuclear Polyhedrosis Virus) Repeat Element Functions as a Powerful Constitutive Enhancer in Transfected Insect Cells

“…They have been identified in all baculoviruses studied thus far, including Bombyx mori NPV (BmNPV), Autographa californica NPV (AcNPV), Choristoneura fumiferana NPV (CfNPV), Lymantria dispar NPV (LdNPV), and Orgyia pseudotsugata NPV (OpNPV) (1)(2)(3)(4)(5)(6)(7)(8). The AcNPV and OpNPV HRs (HR1-HR5) are among the best characterized viral regulatory elements.…”