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发明公开
CN101988049A 共表达小鼠膜型白细胞介素15和Rae-1ε的转基因细胞及其制备方法 失效 - 权利终止

  • 专利标题： 共表达小鼠膜型白细胞介素15和Rae-1ε的转基因细胞及其制备方法
  • 专利标题（英）： Genetically modified cell of coexpression mouse membranous type interleukin 15 and retinoic acid early transcript 1 epsilon (Rae-1 epsilon) and preparation method thereof
  • 申请号： CN201010534375.8
    申请日： 2010-11-08
  • 公开(公告)号： CN101988049A
    公开(公告)日： 2011-03-23
  • 发明人: 钱莉 , 季明春 , 潘兴元 , 龚卫娟 , 田芳 , 段秋芳
  • 申请人： 扬州大学
  • 申请人地址： 江苏省扬州市大学南路88号
  • 专利权人： 扬州大学
  • 当前专利权人： 扬州大学
  • 当前专利权人地址： 江苏省扬州市大学南路88号
  • 代理机构： 南京知识律师事务所
  • 代理商 卢亚丽
  • 主分类号： C12N5/10
  • IPC分类号： C12N5/10 ;  C12N15/12 ;  C12N15/24 ;  C12N15/85
共表达小鼠膜型白细胞介素15和Rae-1ε的转基因细胞及其制备方法
摘要：
本发明涉及涉及一种共表达小鼠膜型白细胞介素15和Rae-1ε的转基因细胞及其制备方法。所说的转基因细胞能稳定共表达膜型IL-15和Rae-1ε蛋白的转基因细胞BaF3细胞。它可通过下述方法制备：利用PCR技术扩增了小鼠Rae-1ε基因和小鼠膜表达型的IL-15基因，将它们分别插入到真核表达载体pVITRO2-mcs的两个多克隆位点，获得重组载体，将重组载体转染小鼠肿瘤细胞株BaF3，经抗生素筛选和流式细胞仪分选后，获得转基因细胞。该转基因细胞可为IKDC提供三重的刺激信号，作为体外高效扩增和活化IKDC的工具。此外，该转基因细胞还可以用于抗肿瘤的免疫治疗研究。
摘要（英）：

The invention relates to a genetically modified cell of coexpression mouse membranous type interleukin 15 and retinoic acid early transcript 1 epsilon (Rae-1 epsilon) and a preparation method thereof. The genetically modified cell is a BaF3 cell which can stabilize the genetically modified cell of the coexpression membranous type IL-15 and Rae-1 epsilon protein. The genetically modified cell can be prepared by a method comprising the following steps: amplifying a mouse Rae-1 epsilon gene and a mouse membrane expression type IL-15 gene by a polymerase chain reaction (PCR) technology; respectively inserting the amplified mouse Rae-1 epsilon gene and the amplified mouse membrane expression type IL-15 gene into two polyclone sites of an eukaryotic expression vector pVITRO2-mcs to acquire a recombined vector; transfecting the recombined vector into a mouse tumor cell strain BaF3; and acquiring the genetically modified cell by antibiotics screening and flow cytometry sorting. The genetically modified cell can provide IFN-gamma producing killer dendritic cells (IKDC) with triple stimulating signals as an instrument for efficiently amplifying and activating the IKDC in vitro. In addition, the genetically modified cell can also be used for antineoplastic immunotherapy research.

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