Construction of an infectious genomic clone of porcine parvovirus: Effect of the 5′-end on DNA replication

Casal, J. Ignacio; Diaz-Aroca, Esmeralda; Ranz, Ana I.; Manclús, Juan J.

doi:10.1016/0042-6822(90)90545-3

Cited by 7 publications

(4 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The overall strategy, similar to that described previously for PPV (Casal et al, 1990), is shown in Fig. 2.…”

Section: Construction Of the Genomic Clonementioning

confidence: 77%

“…The strategy for cloning was similar to that described previously for PPV (Casal et al, 1990)~ A collection of expression clones covering the whole or several regions of the VP2 gene were constructed in the pKK233-2 (Amann & Brosius, 1985), pGEX (Smith & Johnson, 1988) or, by preference, pET vector systems (Studier & Moffatt, 1986). One characteristic of the pET system is the use of Escherichia coli JMI09 (or other similar recA strains) for cloning and maintenance of DNA, after which BL21 is used to express the DNA when the final construct is in the right orientation.…”

Section: Cloning Of the Cp V Genome And Construction Of Recombinant Pmentioning

confidence: 99%

See 1 more Smart Citation

Fine mapping of canine parvovirus B cell epitopes

Turiso¹,

Cortes²,

Ranz³

et al. 1991

Journal of General Virology

View full text Add to dashboard Cite

In this report we describe the topological mapping of neutralizing domains of canine parvovirus (CPV). We obtained 11 CPV-specific monoclonal antibodies (MAbs), six of which are neutralizing. The reactivities were as determined by ELISA and Western blot (immunoblot) analysis. VP2, the most abundant protein of the CPV capsid, seemed to contain all the neutralization sites. Also, an almost full-length genomic clone of CPV was constructed in the bacterial plasmid pUC18 to enable expression of CPV proteins. All the neutralizing MAbs recognized recombinant VP2 when it was expressed as a free protein in Escherichia coli but not when expressed as a fusion protein with glutathione-S-transferase. When two large fragments containing about 85 % and 67 % of the C terminus of VP2 were expressed, no neutralization sites were detected. When fusion proteins containing the N terminus were expressed, two linear determinants were mapped, one between residues 1 to 10 of VP2, and the other between amino acids 11 and 23. The peptide 11 GQPAVRNERATGS 23, recognized by MAb 3C9, was synthesized chemically and checked for immunogenicity, not being able to induce neutralizing activity. Although the antibody response in rabbits to all the fusion proteins was uniformly high, the anti-CPV response was very variable. Protein from pCPVExl 1, which contains a T cell epitope (peptide PKIFINLAKKKKAG) present in the VPl-specific region as well as the B cell epitopes, seemed to be the most effective in inducing virus neutralization.

show abstract

“…The overall strategy, similar to that described previously for PPV (Casal et al, 1990), is shown in Fig. 2.…”

Section: Construction Of the Genomic Clonementioning

confidence: 77%

Section: Cloning Of the Cp V Genome And Construction Of Recombinant Pmentioning

confidence: 99%

Fine mapping of canine parvovirus B cell epitopes

Turiso¹,

Cortes²,

Ranz³

et al. 1991

Journal of General Virology

View full text Add to dashboard Cite

show abstract

“…As the authors speculated [11], shorter and hence packagable transducing genomes were possibly generated from the longer recombinant genome as a result of deletions or illegitimate recombinations. As it has been shown [21,22] deletion mutants of parvovirus genomes can be readily generated from cloned parvovirus genomes transfected into cells. Also it was shown that recombinant parvovirus genomes which were longer than wild type could be associated in vitro with VP in some complexes different from a virion, and delivered by such complexes to new cells [32].…”

Section: Transduction By Jcdnvmentioning

confidence: 98%

“…The integrity of the terminal structures is important because the hairpins serve as primers for synthesis of the second DNA strand during replication of the virus genome. Infectious clones of parvoviruses which have unique sequences at each terminus of the genome (AeDNV for example) must preserve both the left and the right hairpins [21,22]. For parvoviruses with identical sequences at both ends of the genome (AAV2 and JcDNV for example) it is sufficient 3 Fig.…”

Section: Clones Of Densoviruses Are Infectiousmentioning

confidence: 99%

Densovirinae as Gene Transfer Vehicles

Afanasiev

Carlson²

2000

Contributions to Microbiology

View full text Add to dashboard Cite

Densoviruses present an attractive opportunity to develop expression vectors for insects. They exhibit several features that are beneficial for such vectors. The genomes of densoviruses are among the smallest of animal DNA viruses, and, therefore are easier to use for cloning and transfection procedures. The fact that cloned densovirus genomes are infectious greatly simplifies their applications. It seems that both densovirus promoters have high constitutive activity and they can be further transactivated with NS. The major nonstructural protein, NS1, can retain its functions even if fused to a foreign protein. If the recombinant genome carrying a gene of interest is supplied with the missing virus functions, it can be packaged into transducing particles. The packaging capacity of the densovirus particles can accommodate a foreign gene encoding a polypeptide of up to 150 kDa. Finally, the functions missing from the recombinant densovirus genome that are necessary for the production of transducing particles can be supplied in several different ways: they can be expressed from a helper plasmid, in cells transformed with the virus genes, or from an RNA virus. The last two methods produce transducing particles which are free of wild type virus. Densovirus transducing particles can be used for the delivery and expression of a gene of interest in insect cell culture or living insects. Transducing particles expressing reporter genes can be valuable tools in the study of viral pathogenesis and perhaps in the genetic manipulation of insects.

show abstract

Establishment of a rescue system for porcine parvovirus using a seamless cloning method

Zhang

Gao

et al. 2019

Arch Virol

View full text Add to dashboard Cite

Construction of an infectious genomic clone of porcine parvovirus: Effect of the 5′-end on DNA replication

Cited by 7 publications

References 11 publications

Fine mapping of canine parvovirus B cell epitopes

Fine mapping of canine parvovirus B cell epitopes

Densovirinae as Gene Transfer Vehicles

Establishment of a rescue system for porcine parvovirus using a seamless cloning method

Contact Info

Product

Resources

About