We determined the complete nucleotide sequence of bovine parvovirus (BPV), an autonomous parvovirus. The sequence is 5,491 nucleotides long. The terminal regions contain nonidentical imperfect palindromic sequences of 150 and 121 nucleotides. In the plus strand, there are three large open reading frames (left ORF, mid ORF, and right ORF) with coding capacities of 729, 255, and 685 amino acids, respectively. As with all parvoviruses studied to date, the left ORF of BPV codes for the nonstructural protein NS-1 and the right ORF codes for the major parts of the three capsid proteins. The mid ORF probably encodes the major part of the nonstructural protein NP-1. There are promoterlike sequences at map units 4.5, 12.8, and 38.7 and polyadenylation signals at map units 61.6, 64.6, and 98.5. BPV has little DNA homology with the defective parvovirus AAV, with the human autonomous parvovirus B19, or with the other autonomous parvoviruses sequenced (canine parvovirus, feline panleukopenia virus, H-1, and minute virus of mice). Even though the overall DNA homology of BPV with other parvoviruses is low, several small regions of high homology are observed when the amino acid sequences encoded by the left and right ORFs are compared. From these comparisons, it can be shown that the evolutionary relationship among the parvoviruses is B19AAVBPVMVM. The highly conserved amino acid sequences observed among all parvoviruses may be useful in the identification and detection of parvoviruses and in the design of a general parvovirus vaccine. * Corresponding author. of the capsid proteins, whereas the others as a group have significant homology among themselves (5, 48). In this paper, the complete nt sequence of BPV is presented. Genome organization, possible coding regions for the viral proteins, and comparison with other parvoviruses are discussed. The sequence of BPV has several small regions of conservation with other parvoviruses which appear to be useful for understanding the evolutionary relationship of parvoviruses, in the design of vaccines, and for the clinical detection of parvoviruses. Recently the nucleotide sequence of B19, a human autonomous parvovirus, was published (47). The relationship of B19 with other parvoviruses is presented in the section "Comparison with B19" at the end of the Discussion. MATERIALS AND METHODS Materials. Restriction enzymes were purchased from Bethesda Research Laboratories, Inc. (Gaithersburg, Md.), and New England BioLabs, Inc. (Beverly, Mass.). Escherichia coli DNA polymerase I (Klenow fragment) and universal sequencing primer were obtained from Bethesda Research Laboratories. Deoxynucleotides and dideoxynucleotides were purchased from Pharmacia P-L Biochemicals, Inc. (Piscataway, N.J.). Terminal deoxynucleotide transferase, [a.-355]dATP and [a-35S]ddATP were obtained from New England Nuclear Corp. (Boston, Mass.). Various synthetic oligonucleotide primers for DNA-sequencing reactions were the kind gift of R. Foote, Oak Ridge National Laboratory. Cell culture and virus propagation. BPV was p...

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A phosphorylated protein (NP-1) with an Mr of 28,000 has been detected in nuclei of bovine parvovirus (BPV)-infected cells in association with chromatin. No protein in this size range was detected after infection of appropriate cells with several autonomous rodent parvoviruses although the BPV-specific protein is similar in size to noncapsid proteins associated with rabbit parvovirus or adeno-associated virus infection. Structural homology between NP-1 and a BPV capsid protein could be detected by electrophoretic analysis of the products of proteolysis with chymotrypsin. This protein can be detected after in vitro translation of RNA from BPV-infected cells and BPV-specific RNA. Homology between the in vivo-and in vitrosynthesized species was shown by the similarity of the chymotryptic products.

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