Expression of human immunodeficiency virus type 1gag gene using genetically engineered herpes simplex virus type 1 recombinants |
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Authors: | Angela Rösen-Wolff Karl Raab Lothar Zöller Gholamreza Darai Josef Eberle Friedrich Deinhardt |
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Institution: | (1) Institut für Medizinische Virologie der Universität Heidelberg, Heidelberg, FRG;(2) Max von Pettenkofer-Institut für Hygiene und Medizinische Mikrobiologie der Ludwig-Maximilians-Universität München, München, FRG |
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Abstract: | Infectious herpes simplex virus type 1 (HSV-1) recombinants were constructed by inserting the cDNA sequence of the human immunodeficiency virus type 1 (HIV-1)gag gene (from nucleotide position 675 SacI] to 3859 Asp 718] of the cDNA sequences of HIV-1 strain BH-10) within the DNA sequences of theBamHI DNA fragment B of the genome of an apathogenic HSV-1 strain HFEM. This HSV-1 strain possesses a 4.1-kbp deletion within theBamHI DNA fragment B between 0.762 and 0.789 map units of the viral genome, which allows the insertion of at least 4 kbp of foreign genetic material into this particular region. The DNA sequences of the immediate early promoter (IE4) of HSV-1 that were inserted upstream from thegag gene were used as a promoter. The screening of 205 virus stocks derived from individual plaques revealed that 46 recombinant viruses harbor HIV-1gag-specific DNA sequences. However, it was found that only six of the recombinant viruses are able to express thegag gene product of HIV-1. This indicates that the ratio of the positive recombination events is about 2.9%. |
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Keywords: | recombinant plasmid transfection DNA sequencing protein analysis |
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