Epstein-Barr Virus Protocols by Paul J. Farrell (auth.), Joanna B. Wilson, Gerhard H. W. May

By Paul J. Farrell (auth.), Joanna B. Wilson, Gerhard H. W. May (eds.)

The program of recombinant DNA expertise to the research of Epstein-Barr virus (EBV) is swiftly constructing enough perception into the virus-host interplay, in order that its function in affliction pathology is now usually discernible and will more and more be interdicted. In Epstein-Barr Virus Protocols, Joanna Wilson and Gerhard could have assembled a set of the foremost molecular biology protocols utilized in the research of Epstein-Barr virus, in addition to a sequence of necessary immunology, telephone biology, and transgenic mouse protocols. defined in step by step aspect via specialists who use them usually, those with ease reproducible recommendations contain tools for gene expression with mini-EBV plasmids, for expression research by means of FISH, for EBV detection and quantitation, and for phone proliferation and loss of life assays. furthermore, the authors supply info on EBV-based vectors, an updated map of EBV, a finished desk of obtainable latent protein antisera, and assays from in vitro to mobilephone to organ to organism levels.
well timed and hugely functional, Epstein-Barr Virus Protocols presents robust instruments for elucidating the existence cycle of EBV and its host interactions, paintings that provides the emergence of significant new remedies and remedies for EBV-associated illnesses, together with numerous different types of human cancer.

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5. DEPC-treated water. 1% (v/v), stand overnight and then autoclave. 6. Phenol/Chloroform/Isoamyl alcohol (25:24:1). 0, and an equal volume of chloroform containing isoamyl alcohol is added. 7. Absolute ethanol (–20°C). 8. Packaging kits. 9. ). 2. Plating the Library 1. 2. 3. 4. 5. 6. 7. 8. 9. LB broth: 10 g tryptone, 5 g yeast extract, 5 g sodium chloride in 1 L of water. Autoclave. LBM broth: LB broth containing 20 mM magnesium chloride and 2% maltose. 5% agar. Autoclave. 7% agarose. 20% maltose: Filter-sterilized.

Natl. Acad. Sci. USA 74, 1590–1594. 22 Kirchmaier 12. Labarca, C. and Paigen, K. (1980) A simple, rapid, and sensitive DNA assay procedure. Anal. Biochem. 102, 344–352. 13. Mackey, D. and Sugden, B. (1997) Studies on the mechanism of DNA linking by EpsteinBarr virus nuclear antigen 1. J. Biol. Chem. 272, 29873–29879. 14. Graham, F. L. and Van der Eb, A. J. (1973) A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology 52, 456–467. 15. Middleton, T. and Sugden, B. (1992) A chimera of EBNA1 and the estrogen receptor activates transcription but not replication.

4. Chloroform. 5. 100% ethanol. 6. 5 M ammonium acetate. 7. 70% ethanol in H 2O. 8. ). 9. 5 with H2O. 10. Microfuge tubes and microcentrifuge. 2. Agarose Gel Elecrophoresis 1. 2. 3. 4. 5. 6. 1X TBE: 90 mM Tris, 80 mM boric acid, 2 mM EDTA. 5% agarose (as indicated) in 1X TBE, microwaved to melt. 05% Bromophenol Blue, 30% glycerol in H2O. Ethidium bromide (10 mg/mL). Electrophoresis apparatus for slab agarose gels. UV light transiluminator. 3. End-Labeling Primers 1. T4 polynucleotide kinase (New England Biolabs), store at –20°C.

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