ACTIVE GAMMA-CARBOXYLATED HUMAN FACTOR-IX EXPRESSED USING RECOMBINANT DNA TECHNIQUES
ACTIVE GAMMA-CARBOXYLATED HUMAN FACTOR-IX EXPRESSED USING RECOMBINANT DNA TECHNIQUES
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DOI：
10.1038/316268a0
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发表时间：
1985-01-01
期刊：
影响因子：
64.8
通讯作者：
LECOCQ, JP
中科院分区：
文献类型：
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作者：
DELASALLE, H;ALTENBURGER, W;LECOCQ, JP
IX 因子（圣诞因子）是一种在肝脏中产生的维生素 K 依赖性血浆蛋白，在凝血内在途径的中期发挥作用1。凝血因子 IX 的功能缺陷是血友病 B 的基础，血友病 B 是一种 X 染色体连锁隐性疾病，其主要治疗方法是使用凝血因子 IX 浓缩物进行替代治疗。人类因子 IX2-4 基因的克隆和表征意味着通过使用重组 DNA 技术可以以更高的产量和纯度生产人类因子 IX5。我们现在使用人 IX 因子 cDNA 克隆4，将其插入痘苗病毒衍生载体中，以感染通常不产生 IX6 因子的人肝癌细胞和小鼠成纤维细胞。肝癌细胞产生完全活性的因子 IX，而成纤维细胞产生的蛋白质活性低于天然因子 IX，即使存在高水平的维生素 K。人因子 IX 被广泛翻译后修饰，因此可能代表迄今为止通过重组 DNA 技术以活性形式产生的最复杂的蛋白质。我们的研究还说明了基于牛痘病毒的载体除了已被证明可用于生产重组活疫苗之外，还具有在真核细胞中表达大量复杂的、临床上有用的蛋白质的潜力7-9。
Factor IX (Christmas factor), a vitamin K-dependent plasma protein made in the liver, functions in the middle phase of the intrinsic pathway of blood coagulation1. A functional deficiency of factor IX underlies haemophilia B, a chromosome X-linked recessive disease for which the major therapeutic approach is replacement treatment using factor IX concentrates. The cloning and characterization of the gene for human factor IX2–4would mean that human factor IX could be produced in greater yield and purity through using recombinant DNA techniques5. We have now used a human factor IX cDNA clone4, inserted into a vaccinia virus-derived vector, to infect human hepatoma cells which normally produce no factor IX6, and mouse fibroblasts. Fully active factor IX was produced by the hepatoma cells, whereas the fibroblasts produced a protein less active than natural factor IX, even in the presence of high levels of vitamin K. Human factor IX is extensively post-translationally modified, and thus represents probably the most complex protein produced in active form by recombinant DNA techniques to date. Our study also illustrates the potential of vaccinia virus-based vectors for expressing significant amounts of complex, clinically useful proteins in eukaryotic cells, in addition to its already demonstrated usefulness for producing live recombinant vaccines7–9.