公开(公告)号：US5681733A

公开(公告)日：1997-10-28

申请号：US258639

申请日：1994-06-10

申请人： Hongsheng Su ,  Francoise Blain ,  Clark Bennett ,  Kangfu Gu ,  Joseph Zimmermann ,  Roy Musil

发明人： Hongsheng Su ,  Francoise Blain ,  Clark Bennett ,  Kangfu Gu ,  Joseph Zimmermann ,  Roy Musil

IPC分类号： C12N15/09 ,  C07K16/40 ,  C12N1/21 ,  C12N9/88 ,  C12N15/60 ,  C12N15/74 ,  C12P21/08 ,  C12R1/19

CPC分类号： C12Y402/02007 ,  C12N15/74 ,  C12N9/88

摘要： The present invention describes the isolation and sequence of genes from Flavobacterium heparinum encoding heparin and heparan sulfate degrading enzymes, heparinase II and heparinase III (EC 4.2.2.8). It further describes a method of expressing and an expression for heparinases I, II and III using a modified ribosome binding region derived from a promoter from glycosaminoglycan lyase genes of F. heparinum. Also, a multi-step protein purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography and hydroxylapatite chromatography is outlined. Antibodies against a post-translational modification moiety common to Flavobacterium heparinum proteins and a method to obtain antibodies specific to these moieties and to the amino acid sequences of heparinases I, II and III are described.

摘要翻译： 本发明描述了编码肝素黄肝素和硫酸乙酰肝素降解酶，肝素酶II和肝素酶III（EC 4.2.2.8）的基因的分离和序列。 它进一步描述了使用衍生自肝素肝素的糖胺聚糖裂解酶基因的启动子的修饰的核糖体结合区域来表达和表达肝素酶I，II和III的方法。 此外，概述了掺入细胞破碎，阳离子交换层析，亲和层析和羟基磷灰石层析的多步蛋白质纯化方法。 描述了针对肝炎黄杆菌蛋白质共同的翻译后修饰部分的抗体以及获得对这些部分和肝素酶I，II和III的氨基酸序列特异性的抗体的方法。

公开(公告)号：US5919693A

公开(公告)日：1999-07-06

申请号：US900951

申请日：1997-07-25

申请人： Hongsheng Su ,  Francoise Blain ,  Clark Bennett ,  Kangfu Gu ,  Joseph Zimmermann ,  Roy Musil

发明人： Hongsheng Su ,  Francoise Blain ,  Clark Bennett ,  Kangfu Gu ,  Joseph Zimmermann ,  Roy Musil

IPC分类号： C12N15/09 ,  C07K16/40 ,  C12N1/21 ,  C12N9/88 ,  C12N15/60 ,  C12N15/74 ,  C12P21/08 ,  C12R1/19 ,  C12N15/63

CPC分类号： C12Y402/02007 ,  C12N15/74 ,  C12N9/88

摘要： The present invention describes the isolation and sequence of genes from Flavobacterium heparinum encoding heparin and heparan sulfate degrading enzymes, heparinase II and heparinase III (EC 4.2.2.8). It further describes a method of expressing and an expression for heparinases I, II and III using a modified ribosome binding region derived from a promoter from glycosaminoglycan lyase genes of F. heparinum. Also, a multi-step protein purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography and hydroxylapatite chromatography is outlined. Antibodies against a post-translational modification moiety common to Flavobacterium heparinum proteins and a method to obtain antibodies specific to these moieties and to the amino acid sequences of heparinases I, II and III are described.

摘要翻译： 本发明描述了编码肝素黄肝素和硫酸乙酰肝素降解酶，肝素酶II和肝素酶III（EC 4.2.2.8）的基因的分离和序列。 它进一步描述了使用衍生自肝素肝素的糖胺聚糖裂解酶基因的启动子的修饰的核糖体结合区域来表达和表达肝素酶I，II和III的方法。 此外，概述了掺入细胞破碎，阳离子交换层析，亲和层析和羟基磷灰石层析的多步蛋白质纯化方法。 描述了针对肝炎黄杆菌蛋白质共同的翻译后修饰部分的抗体以及获得对这些部分和肝素酶I，II和III的氨基酸序列特异性的抗体的方法。