Molecular cloning and clones of human b lymphotropic virus...

C - Chemistry – Metallurgy – 12 – N

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195/33, 195/1.22

C12N 15/86 (2006.01) C07K 14/03 (2006.01) C12N 7/00 (2006.01) C12N 7/01 (2006.01) C12N 7/02 (2006.01) C12Q 1/70 (2006.01)

Patent

CA 1322342

ABSTRACT OF THE DISCLOSURE A clone of Human B Lymphotropic Virus (HBLV), pZVH14 has been isolated (from the peripheral blood leukocytes of six individuals) and characterized. The clone is obtained from nucleic acids extracted from purified virus, and contains a 9.0 Kb HindIII fragment of HBLV . HBLV contains a double-stranded DNA genome and is morphologically similar to some members of the Herpes family. The clone is lymphotropic and selectively infects freshly isolated human B cells, where it induces the appearance of characteristic large refractile mono- nucleated or binucleated host cells. The molecular clone of the present invention is capable of in situ hybridization with host cells infected with HBLV. This clone, therefore, is useful as a diagnostic probe for HBLV-infected cells, as well as for the detection of viral antigens or antibodies in blood samples (using an immunofluorescence assay or any other assay for antigens or antibodies which uses viral nucleic acids). Molecular clone pZVH14 is also used in the detection of early virus infection of umbilical cord blood lymphocytes and spleen cells by in situ hybridization in culture. The present invention is also capable of introducing genes into target cells of the virus as a recombinant DNA virus vector system.

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