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Recombinant fungi strain and method for production of human lactoferrin antibacterial polypeptide derived from human thereof

机译:重组真菌菌株及其衍生自人的人乳铁蛋白抗菌多肽的生产方法

摘要

PURPOSE: Provided are recombinant fungus strains which express human lactoferrin and lactoferrin antibacterial polypeptides in a high yield and is thus useful for manufacturing medicines and protecting livestock from various diseases. CONSTITUTION: The recombinant fungus strains MMS-1 (KCTC 0683BP) and MLS60 (KCTC 0684BP) are obtained by the next steps of: i) selecting and cultivating strain RIB40 unable to generate arginine dehydrogenase; ii) multiplying human lactoferrin and its antibacterial polypeptides through PCR; iii) cutting cDNAs of the human lactoferrin and its antibacterial polypeptides and ligating changed cDNAs of the human lactoferrin and its antibacterial polypeptides with vectors pFGC33 and pTAex3 respectively; iv) transforming strain DH5- with the two vectors; v) separating recombinant vectors pFGHLF and pGLC from the transformants; vi) inserting argB gene into vectors pTAex3 and pUC118 and transforming E. coli with the vectors to obtain selection markers pDEargB and pMAR1; and vii) transforming strain RIB40 with the recombinant vector pFGHLF and the selection marker pDEargB and with the vector pGLC and the selection marker pMAR1. Human lactoferrin and human lactoferrin antibacterial polypeptide are prepared by fermenting culturing each strain MMS-1 and MLS60.
机译:目的:提供重组真菌菌株,其以高产量表达人乳铁蛋白和乳铁蛋白抗菌多肽,因此可用于制造药物和保护牲畜免受各种疾病侵害。组成:重组真菌菌株MMS-1(KCTC 0683BP)和MLS60(KCTC 0684BP)是通过以下步骤获得的:i)选择和培养不能产生精氨酸脱氢酶的RIB40菌株; ii)通过PCR扩增人乳铁蛋白及其抗菌多肽; iii)切割人乳铁蛋白及其抗菌多肽的cDNA,并将人乳铁蛋白及其抗菌多肽的改变的cDNA分别与载体pFGC33和pTAex3连接。 iv)用两个载体转化DH5-菌株; v)从转化体中分离重组载体pFGHLF和pGLC; vi)将argB基因插入载体pTAex3和pUC118中,并用该载体转化大肠杆菌,获得选择标记pDEargB和pMAR1。 vii)用重组载体pFGHLF和选择标记pDEargB以及载体pGLC和选择标记pMAR1转化RIB40菌株。通过发酵培养每种菌株MMS-1和MLS60来制备人乳铁蛋白和人乳铁蛋白抗菌多肽。

著录项

  • 公开/公告号KR100383436B1

    专利类型

  • 公开/公告日2003-05-12

    原文格式PDF

  • 申请/专利权人

    申请/专利号KR19990050314

  • 发明设计人 성창근;문태현;민대규;

    申请日1999-11-12

  • 分类号C12N15/12;C12N15/63;C12N1/20;

  • 国家 KR

  • 入库时间 2022-08-21 23:45:26

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