FIELD: genetic engineering. SUBSTANCE: rabies virus (isolate "Vnukovo-32") was reproduced in cell culture of kidneys of golden hamster. Virions in cultural fluid were concentrated by means of ultrafiltration through special filters and then settled by ultracentrifugation. Virional RNA was separated by means of guanidine-thiocyanate and used for synthesis of cDNA gene of glycoprotein G. cDNA was amplified by means of specific primers according to method of polymerase chain reaction (PCR). Amplified gene of glycoprotein G was cloned in cells E. coli by means of procaryotic vector pUC-19. Sequence of nucleotides and amino acids was detected according to method of Senger. Gene of glucoprotein G was recloned into expressing procaryotic vector pUC-18. Expressed viral protein showed specific immunogenic activity. EFFECT: possibility of use in construction of vaccines against rabies and diagnostic preparations.
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Authors
Dates
1994-02-28—Published
1991-12-18—Filed