首页> 外国专利> PRODUCTION METHOD OF RECOMBINANT HUMAN LACTOFERRIN, N-LOBE LACTOFERRIN, AND C-LOBE LACTOFERRIN USING PICHIA PASTORIS LF2 (KFCC 11106), PICHIA PASTORIS NFL3 (KFCC 11107), AND PICHIA PASTORIS CFL3 (KFCC 11105)

PRODUCTION METHOD OF RECOMBINANT HUMAN LACTOFERRIN, N-LOBE LACTOFERRIN, AND C-LOBE LACTOFERRIN USING PICHIA PASTORIS LF2 (KFCC 11106), PICHIA PASTORIS NFL3 (KFCC 11107), AND PICHIA PASTORIS CFL3 (KFCC 11105)

机译：使用PICHIA PASTORIS LF2（KFCC 11106），PICHIA PASTORIS NFL3（KFCC 11107）和PICHIA PASTORIS CFL3（KFCC 11105）生产重组人乳铁蛋白，N-叶乳铁蛋白和C-叶乳铁蛋白的方法

页面导航

摘要
著录项
相似文献

摘要

PURPOSE: A concentration of lactoferrin which is physiologically activating protein, in the milk is low compared with in fore milk, so addition of lactoferrin is required. Lactoferrin having antibiotic activity and regulation of iron uptake in the intestine of farm animals is an important ingredient of animal feed stuff. A production method of lactoferrin using Pichia pastor is provided. CONSTITUTION: cDNAs for lactoferrin, N-lobe lactoferrin, and C-lobe lactoferrin are amplified by PCR. Amplified PCR products are modified by restriction of signal sequence and then ligated with pPIC9K vector. The vector is introduced into E. coli Top10F' strain and transforms it. Each recombinant vector, purified from E. coli is introduced into histidine auxotroph Pichia pastor is and transforms it. Correct colonies are selected. Second round selection is performed on the ground of kanamycin resistance. Using SDS-PAGE and PCR amplification, expression of each protein is confirmed. Preinduction with glycerol and induction with methanol are used to produce each protein. After the main culture, the culture broth and cells are freeze dried and mixed in the animal feed stuff.

机译：目的：乳汁中的生理活化蛋白乳铁蛋白的浓度低于原乳，因此需要添加乳铁蛋白。在农场动物的肠道中具有抗生素活性和铁吸收调节作用的乳铁蛋白是动物饲料的重要成分。提供了一种使用毕赤氏酵母生产乳铁蛋白的方法。组成：乳铁蛋白，N-叶乳铁蛋白和C-叶乳铁蛋白的cDNA通过PCR扩增。通过限制信号序列来修饰扩增的PCR产物，然后与pPIC9K载体连接。将该载体导入大肠杆菌Top10F'菌株并转化。从大肠杆菌纯化的每个重组载体都被引入到组氨酸营养缺陷型毕赤酵母中并转化。选择正确的菌落。基于卡那霉素抗性进行第二轮选择。使用SDS-PAGE和PCR扩增，可以确认每种蛋白质的表达。用甘油预诱导和用甲醇诱导产生每种蛋白质。主要培养后，将培养液和细胞冷冻干燥并在动物饲料中混合。

著录项

公开/公告号KR20000029179A

专利类型
公开/公告日2000-05-25

原文格式PDF
申请/专利权人 EASY BIO SYSTEM INC.;
展开▼

申请/专利号KR19990045290
发明设计人 BOK JIN DEOK;SEONG CHANG GEUN;SHIN MIN SEON;MUN TAE HYUN;CHOI YOON JAE;YANG JAE CHEON;
展开▼

申请日1999-10-19
分类号C12N15/63;C12N15/12;C12N15/81;
国家 KR
入库时间 2022-08-22 01:45:50

相似文献

专利
外文文献
中文文献