首页> 外国专利> RECOMBINANT PLASMID DNA pAK380 CODING POLYPEPTIDE OF RECOMBINANT FACTOR IX OF HUMAN BLOOD COAGULABILITY, LINE OF CELLS Cricetulus griseus CHO, 1E6-PRODUCER OF RECOMBINANT FACTOR IX OF HUMAN BLOOD COAGULABILITY, AND METHOD FOR OBTAINING POLYPEPTIDE HAVING ACTIVITY OF RECOMBINANT FACTOR IX

RECOMBINANT PLASMID DNA pAK380 CODING POLYPEPTIDE OF RECOMBINANT FACTOR IX OF HUMAN BLOOD COAGULABILITY, LINE OF CELLS Cricetulus griseus CHO, 1E6-PRODUCER OF RECOMBINANT FACTOR IX OF HUMAN BLOOD COAGULABILITY, AND METHOD FOR OBTAINING POLYPEPTIDE HAVING ACTIVITY OF RECOMBINANT FACTOR IX

机译：重组人血浆凝血因子IX的质粒pAK380编码多肽，灰线虫CHO细胞株，人血凝固性IX因子的1E6-生产者及结合力的研究方法

页面导航

摘要
著录项
相似文献

摘要

FIELD: biotechnologies.;SUBSTANCE: invention can be used for obtaining recombinant blood coagulability factor IX of human being (hFIX). Recombinant plasmid DNA pAK380 containing gene of protein rhFIX, MAR - binding area to nuclear matrix of lysozyme gene of birds, virus transcription enhancer CMV and an internal translation initiation site IRES of encephalomyocarditis virus, gene DHFR of a mouse, a polyadenylation signal of virus SV40, gene of aminoglycoside-3'-phosphotransferase for stability to geneticin (Neo), a cassette for expression in bacteria cells of gene β-lactamase for stability to ampicillin, is used for obtaining recombinant factor hFIX in cells of line Cricetulus griseus CHO 1E6. By transformation of cell line C. griseus CHO DHFR - recombinant plasmid DNA pAK380 there obtained is cell line C. griseus CHO 1E6 producing recombinant hFIX with stable high yield at the level of 50 mg/l/24 h. After cultivation of cells-producers there extracted is hFIX by pseudoaffine chromatography on Q Sepharose with elution of 10mM CaCl2; then, on Heparin-Sepharose FF with elution of 600 mM NaCl, and chromatography on hydroxyapatite of type I with elution of 600 mM K3PO3 and chromatography on Source 30Q with elution of 600 mM with ammonium acetate.;EFFECT: improvement of the method.;4 cl, 5 dwg, 7 ex, 3 tbl

机译：领域：本发明可用于获得人的重组凝血因子IX（hFIX）。重组质粒DNA pAK380，包含蛋白rhFIX基因，MAR-鸟类溶菌酶基因核基质的结合区，病毒转录增强剂CMV和脑心肌炎病毒的内部翻译起始位点IRES，小鼠的基因DHFR，病毒SV40的多腺苷酸化信号用于对遗传霉素稳定的氨基糖苷3'-磷酸转移酶（Neo）基因是一种在细菌细胞中表达对氨苄青霉素稳定的表达盒，用于在灰ric（Cricetulus griseus）CHO 1E6系的细胞中获得重组因子hFIX。通过转化细胞系灰霉菌CHO DHFR-重组质粒DNA pAK380，获得了细胞系灰霉菌CHO 1E6，其以50mg / l / 24h的水平稳定地高产量产生重组hFIX。培养细胞产生物后，通过假石蜡层析在Q琼脂糖凝胶上用10mM CaCl 2洗脱提取hFIX。然后，在肝素-琼脂糖FF上用600 mM NaCl洗脱，在I型羟基磷灰石上用600 mM K3PO3洗脱，并在Source 30Q上用600 mM乙酸铵洗脱。 4厘升，5载重吨，7吨，3吨

著录项

公开/公告号RU2500816C1

专利类型
公开/公告日2013-12-10

原文格式PDF
申请/专利权人 OBSHCHESTVO S OGRANICHENNOJ OTVETSTVENNOSTJU MEZHDUNARODNYJ BIOTEKHNOLOGICHESKIJ TSENTR GENERIUM (MBTS GENERIUM);
展开▼

申请/专利号RU20120130854
发明设计人 GRIGOREVA OLGA VASILEVNA;ZAVALNYJ MIKHAIL ALEKSANDROVICH;KARPOV ANDREJ PAVLOVICH;PETROV ANDREJ VLADIMIROVICH;FABRICHNYJ IGOR PAVLOVICH;SHUSTER ALEKSANDR MIKHAJLOVICH;
展开▼

申请日2012-07-19
分类号C12N15/11;C12N15/63;C12N5/10;C12P21/02;
国家 RU
入库时间 2022-08-21 15:38:48

相似文献

专利
外文文献