RDNA NTS BASED GENE MULTIPLE INSERTION CASSETTE SET AND GRAS-GRADE RECOMBINANT YEAST STRAIN
    6.
    发明公开
    RDNA NTS BASED GENE MULTIPLE INSERTION CASSETTE SET AND GRAS-GRADE RECOMBINANT YEAST STRAIN 审中-公开
    基于RDNA NTS的基因多重插入片段集和GRAS-GRADE重组酵母菌株

    公开(公告)号:EP3184639A1

    公开(公告)日:2017-06-28

    申请号:EP14900237.0

    申请日:2014-12-30

    摘要: The present invention relates to a ribosomal DNA nontranscribed spacer (rDNA NTS)-based gene multiple insertion cassette set and a GRAS-grade recombinant yeast strain using the same. In particular, a gene multiple insertion cassette set including rDNA NTS fragments and an auxotrophic selection marker having an incomplete promoter is developed, and a safe oral recombinant strain having no antibiotic resistant marker is constructed by multiple insertion of an optimum number of the developed gene multiple insertion cassette sets into chromosomes of a Saccharomyces cerevisiae strain. In addition, the present invention provides a vaccine composition including, as an active ingredient, the above strain, a culture product thereof, a cell lysate, or nodavirus capsid protein (NNVcp) isolated and purified therefrom. In addition, the present invention provides a composition for feed addition including, as an active ingredient, the above strain, a culture product thereof, a cell lysate, or squalene or oxidosqualene isolated and purified therefrom. Furthermore, the present invention provides a technique of preparing a GRAS-grade recombinant yeast strain for mass production of various recombinant proteins or useful metabolites.

    摘要翻译: 本发明涉及基于核糖体DNA非转录间隔区(rDNA NTS)的基因多重插入盒组和使用其的GRAS级重组酵母株。 特别地,开发了包含rDNA NTS片段的基因多重插入盒组和具有不完全启动子的营养缺陷型选择标记,并且通过多次插入最适数量的显现基因倍数构建不具有抗生素抗性标记的安全口服重组菌株 插入盒置于酿酒酵母菌株的染色体中。 此外,本发明提供了一种疫苗组合物,其包含从其分离和纯化的上述菌株,其培养产物,细胞裂解物或诺达病毒衣壳蛋白(NNVcp)作为活性成分。 此外,本发明提供了一种用于饲料添加的组合物,其包含作为活性成分的上述菌株,其培养产物,细胞裂解物或由其分离和纯化的角鲨烯或氧化角鲨烯。 此外,本发明提供了制备用于大量生产各种重组蛋白质或有用代谢物的GRAS级重组酵母菌株的技术。