首页> 外国专利> METHYLOTROPHIC YEAST STRAIN PICHIA PASTORIS YST-PPIC9PGAPZALPHA-SHORT_HTFNG, PRODUCING RECOMBINANT HUMAN TRANSFERRIN CONTAINING AMINO ACID SUBSTITUTES N413D AND N611D

METHYLOTROPHIC YEAST STRAIN PICHIA PASTORIS YST-PPIC9PGAPZALPHA-SHORT_HTFNG, PRODUCING RECOMBINANT HUMAN TRANSFERRIN CONTAINING AMINO ACID SUBSTITUTES N413D AND N611D

机译:甲基营养酵母菌株Pichia Pastoris YST-PPIC9PGAPZALPHA-SHORT_HTFNG,生产包含氨基酸的重组人转铁蛋白,取代了N413D和N611D

摘要

FIELD: biotechnology.;SUBSTANCE: invention relates to biotechnology, specifically to production of modified recombinant human transferrin containing amino acid substitutes N413D and N611D (TGF). Invention can be used to produce serum-free nutrient media for culturing mammalian cells. Invention is produced on the basis of GS115 strain of methylotrophic yeast P. pastoris strain-producer Yst-pPIC9pGAPZalpha-short_hTFNG, containing in its genome integrated expression genetic construct pPIC9GAPalpha-short_hTFNG and pGAPZalpha-short_hTFNG, each of which contains a cDNA sequence coding a pre-sequence of the alpha-short signal peptide, which enables to release TGF into the culture medium, in single reading frame with nucleotide sequence of cDNA of modified human serum human transferrin (N413Q and N611Q) gene.;EFFECT: invention enables to obtain recombinant human serum transferrin, which does not contain N-glycosylation, in a eukaryotic system of expression of methylotrophic yeast P_ pastoris with a product level of 100 mg/l in growth conditions in a minimum salt medium in a fermenter.;1 cl, 3 dwg, 3 ex
机译:技术领域本发明涉及生物技术,尤其涉及包含氨基酸替代物N413D和N611D(TGF)的修饰的重组人转铁蛋白的生产。本发明可用于生产用于培养哺乳动物细胞的无血清营养培养基。基于甲基营养酵母巴斯德毕赤酵母菌株生产者Yst-pPIC9pGAPZalpha-short_hTFNG的GS115菌株,在其基因组中包含整合表达基因构建体pPIC9GAPalpha-short_hTFNG和pGAPZalpha-short_hTFNG,它们各自包含编码前体的cDNA序列。 -短信号肽序列,该序列能够在修饰的人血清人转铁蛋白(N413Q和N611Q)基因的cDNA的核苷酸序列的单一阅读框中将TGF释放到培养基中。在发酵罐的最低盐培养基中,在生长条件下,在生长条件下以100 mg / l的水平生产甲基营养酵母P_pastoris的真核表达系统中不含N-糖基化的人血清转铁蛋白.1 cl,3 dwg ,3前

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