 Herpes treatment |
| I claim: 1. A method for the control of herpes virus infections in man which comprises contacting ... |
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| Dense star polymers and dendrimers |
| OF ILLUSTRATIVE EMBODIMENTS In the dense star polymers of the present invention, the core is ... |
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| Glycerin derivatives and inhibition of blood PAF |
| OF THE INVENTION The glycerine derivatives of the invention are compounds of the formula ##STR3## ... |
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| Oligonucleotide derivatives and production thereof |
| Gist In view of the state of the art as described above, the present inventors have developed an ... |
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| Process for synthesizing macrocyclic chelates |
| The invention is a process for synthesizing a 12 membered ring tetraaza macromolecule. The process ... |
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| Immunoconjugates joined by thioether bonds having reduced toxicity and improved selectivity |
| I claim: 1. A method for producing a 1:1 protein:anitbody tioether-linked immunoconjugate as a ... |
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| Compositions for photodynamic therapy |
| I claim: 1. A pharmaceutical composition useful for photodynamic therapy which composition contains,... |
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| Amplifier molecules for enhancement of diagnosis and therapy |
| As used herein, an "amplifier" or "amplifier molecule" is a chemical compound having a ... |
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| Peptide-containing polyethylene glycol derivatives and application thereof |
| OF THE PREFERRED EMBODIMENTS The present invention will hereinafter be explained in more detail. T... |
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| Modified HIL-6 |
| OF THE INVENTION Described below is the chemical modification with PEG of at least one amino group ... |
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Multigene vectors
| Details |
Inventors: Glorioso, Joseph C.; Krisky, David;
Assignee: University of Pittsburgh of the Commonwealth System of Higher Education (Pittsburgh, PA)
Primary Examiner: Brusca; John S.
Assistant Examiner:
Attorney, Agent or Firm: Leydig, Voit & Mayer, Ltd.
The present invention provides a method for preparing HSV vectors. The method comprises co-transfecting a source vector and a mutating cassette together into a population of appropriate host cells, such that homologous recombination occurs between the mutating cassette and the source vector whereby the mutating cassette replaces a region of the HSV genome. The mutating cassette has a unique restriction site not present in the sequence of the vector. The method further comprises plaquing the co-transfected host cells, selecting plaques in which recombination has occurred between the source vector and the mutating cassette, and isolating the viral DNA from the plaques. The isolated viral DNA is digested with a restriction endonuclease appropriate for cleaving the viral DNA at the unique restriction site within the mutating cassette to produce two viral polynucleotides. Following purification, the two viral polynucleotides can be ligated to form an HSV vector comprising the two viral polynucleotides. Alternatively, the two isolated viral polynucleotides can be recombined with an insertion cassette to form an HSV vector comprising the insertion cassette at the former locus of the unique restriction site. The present invention further provides a mutant vector, particularly an HSV vector constructed in accordance with the method for the present invention. The present invention further provides a multigene HSV vector, particularly a multigene HSV vector for cancer therapy. |
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DETAILED DESCRIPTION The present invention provides a method for preparing HSV vectors.
The method comprises co-transfecting a source vector and a mutating cassette together into a population of appropriate host cells, such that homologous recombination occurs between the mutating cassette and the source vector whereby the mutating cassette replaces a region of the HSV genome.
The mutating cassette has a unique restriction site not present in the sequence of the vector.
The method further comprises plaquing the co-transfected host cells, selecting plaques in which recombination has occurred between the source vector and the mutating cassette, and isolating the viral DNA from the plaques.
The isolated viral DNA is digested with a restriction endonuclease appropriate for cleaving the viral DNA at the unique restriction site within the mutating cassette to produce two viral polynucleotides.
Following purification, the two viral polynucleotides can be joined to form an HSV vector comprising the two viral polynucleotides.
Alternatively, the two isolated viral polynucleotides can be joined with an insertion cassette to form an HSV vector comprising the insertion cassette at the former locus of the unique restriction site.
The present invention further provides a mutant vector, particularly an HSV vector constructed in accordance with the method for the present invention.
The present invention further provides a multigene HSV vector, particularly a multigene HSV vector for cancer therapy.
The present invention will prove highly useful in biological research.
Specifically, the present invention provides reagents and methods enabling biologists to more easily study HSV molecular genetics and cytotoxicity.
Additionally, the present invention provides reagents and methods permitting biologists to investigate the cell biology of viral growth and infection.
Furthermore, the multigene vectors of the present invention will equip the biologist with novel tools for investigating molecular and cellular biology of gene expression and regulation in novel genetic backgrounds
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Gene Therapy 
