公开(公告)号：US5854031A

公开(公告)日：1998-12-29

申请号：US748947

申请日：1996-11-14

申请人： Yoshifumi Jigami ,  Ken-ichi Nakayama ,  Yoh-ichi Shimma ,  Xiao-hui Wang

发明人： Yoshifumi Jigami ,  Ken-ichi Nakayama ,  Yoh-ichi Shimma ,  Xiao-hui Wang

IPC分类号： C12N15/09 ,  C07H21/04 ,  C12N1/19 ,  C12N9/12 ,  C12P19/04 ,  C12P19/26 ,  C12R1/865 ,  C12N9/10 ,  C12N15/63

CPC分类号： C12N9/1205 ,  C12P19/04

摘要： The present invention relates to a mannose-1-phosphate transferase gene from yeast (Saccharomyces cerevisiae), a process for producing a mannose-1-phosphate-containing acidic sugar chain which comprises culturing yeast cells transformed with a plasmid DNA containing said gene, obtaining mannose-1-phosphate transferase from the culture, and allowing the enzyme to act on a neutral core sugar chain in vivo or in vitro, and a process for producing a phosphate-containing acidic sugar chain which comprises removal of a mannose moiety by acid-treatment of said mannose-1-phosphate-containing acidic sugar chain. According to the present invention, an acidic sugar chain having mannose-1-phosphate added to the same neutral core sugar chain as a high mannose type sugar chain produced by mammalian (e.g. human) cells can be produced in large amounts with high purity by genetic engineering means using yeast. Further, an acidic sugar chain identical with a useful phosphate-containing core sugar chain derived from human, which can function as a labeling marker for transport of glycoproteins into lysosomes in mammalian (e.g. human) cells, can be produced in large amounts with high purity.

摘要翻译： 本发明涉及来自酵母（酿酒酵母）的甘露糖-1-磷酸转移酶基因，含有甘露糖-1-磷酸的酸性糖链的制备方法，其包括培养用含有该基因的质粒DNA转化的酵母细胞，得到 来自培养物的甘露糖-1-磷酸转移酶，并且允许酶在体内或体外作用于中性核心糖链，以及生产含磷酸盐酸性糖链的方法，其包括通过酸 - 磷酸转移酶去除甘露糖部分， 处理所述含甘露糖-1-磷酸的酸性糖链。 根据本发明，通过哺乳动物（例如人）细胞产生的与甘露糖型糖链相同的中性核心糖链上添加的具有甘露糖-1-磷酸的酸性糖链可通过遗传学以高纯度大量生产 工程意味着使用酵母。 此外，可以以高纯度大量生产与用于将哺乳动物（例如人）细胞中的糖蛋白转运到溶酶体中的可用作标记标记的来源于人的有用的含磷酸核心糖链相同的酸性糖链 。