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METHOD FOR CREATING TRANSGENIC ANIMALS WITH STABLE AND HIGH TARGET PROTEIN EXPRESSION IN MILK

机译:稳定,高目标蛋白表达的奶中转基因动物的培养方法

摘要

FIELD: medicine.;SUBSTANCE: invention refers to biotechnology, particularly to methods for preparing next generation drug preparations and biologically active additives in bioreactors on the basis of transgenic producing mammals. The method for creating transgenic animals producing a protein with stable and high expression in milk, involves producing transgenic mammals with using a vector containing a reporter gene coding a target protein that is a goat beta-casein gene promoter, a bovine growth factor terminator and effective two-fold transcription terminators. The terminators surround an expression cartridge and possess an ability to break genome transcripts effectively in a mammalian genome by the effective protection of transgene expression in the mammalian genome against further repression. The effective two-fold terminators represent any mammalian genome site fulfilling the following conditions: 3'-sites of the two simultaneously expressing and opposite genes containing a site of the second-to-last exon, the last intervening sequence, the last exon and a polyadenylation signal, a space of two polyadenylation signals at different DNA strands is no more than 100 base pairs.;EFFECT: method can be used for creating the transgenic animals with high and stable target protein production in milk for medical and research purposes.;4 dwg, 4 tbl, 3 ex
机译:技术领域本发明涉及生物技术,尤其涉及基于转基因生产的哺乳动物在生物反应器中制备下一代药物制剂和生物活性添加剂的方法。产生转基因动物的方法,所述转基因动物产生在牛奶中稳定和高表达的蛋白质,包括使用含有编码靶蛋白的报告基因的载体来产生转基因哺乳动物,所述靶蛋白是山羊β-酪蛋白基因启动子,牛生长因子终止子且有效双重转录终止子。终止子围绕表达盒,并具有通过有效保护哺乳动物基因组中的转基因表达免受进一步阻遏而有效破坏哺乳动物基因组中的基因组转录物的能力。有效的双重终止子代表满足以下条件的任何哺乳动物基因组位点:两个同时表达且相对的基因的3'位点,包含倒数第二个外显子,最后一个插入序列,最后一个外显子和一个聚腺苷酸化信号,在不同的DNA链上两个聚腺苷酸化信号的间隔不超过100个碱基对。;效果:该方法可用于创建具有高且稳定的目标蛋白产量的转基因动物,用于医学和研究目的; 4 dwg,4汤匙,3前

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