HYBRID GENES BLG-HIFN-G BASED ON REGULATORY SEQUENCE OF SHEEP BETA-LACTOGLOBULIN GENOME GENE FOR EXPRESSION OF HUMAN GAMMA-INTERFERON IN TRANSGENIC ANIMAL MAMMA GLAND

摘要

FIELD: molecular biology, genetic engineering. SUBSTANCE: hybrid genes BLG-hIFN-g2 and BLG-hIFN-g3 have length about 10 thousand base pairs (t.b.p. ) and consist of: Sal I-SnaB I-fragment (length is 4.7 t.b.p.) that is 5'-flanking sequence of sheep beta-globulin gene (BLG); SnaB I-Asu II-fragment (1.7 t. b. p. ) that is part of human gamma-interferon genome gene (hIFN-g) involving the first exon without the first 113 b. p. of its nontranslatable region, the first two introns, the first two and part of the third exon; Asu II-Bam HI-fragment that is in the case of BLG-hIFN-g2 gene a sequence involving part of the third and all fourth exon, the third intron and part of 3'-flanking sequence of gene hIFN-g; in the case of gene BLG-hIFN-g3 - the sequence involving part of the third and all fourth exon of gene hIFN-g and part of the sixth exon, the sixth intron, the seventh exon containing polyadenylation signal, part of 3'-flanking sequence, part of the first, the second and third exons, two the first and part of the third intron of gene BLG; Bam HI-Sal I-fragment that is part of polylinker of plasmid pUC18; hybrid genes contain: unique recognition site for restriction endonuclease Sna BI artificially inserted into site Pvu II of nontranslatable part of the first exon of gene BLG. Genes BLG-hIFN-g provide expression of hIFN-g in mamma gland of transgenic animals. Milk of transgenic mouse females shows interferon antiviral activity 10⁶-10⁷ IU/ml for hybrid gene BLG-hIFN-g2 and 6000-40000 IU/ml for hybrid gene BLG-hIFN-g3. EFFECT: design of hybrid genes indicated above.