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Bioassay of the soluble human tumor necrosis factor receptor Ⅰrecombinant plasmid expression in vitro
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- Author:
- No author available
- Journal Title:
- CHINESE JOURNAL OF STOMATOLOGY
- Issue:
- 3
- DOI:
- 10.3760/j.issn:1002-0098.2004.03.003
- Key Word:
- 受体,肿瘤坏死因子;质粒;转染;CHO细胞
Abstract： 目的检测构建的PcDNA3.1-sTNFRⅠ真核表达载体在体外转染哺乳动物细胞后能否有效表达目的产物,表达的目的产物是否具有生物学活性,为探索sTNFRⅠ基因治疗牙周炎奠定一定的实验基础.方法 ELISA法检测PcDNA3.1-sTNFRⅠ转染的CHO细胞培养上清液中sTNFRⅠ的表达,MTT法检测表达产物是否具有抑制肿瘤坏死因子α( TNF-α)对L929的细胞毒作用的能力.结果重组质粒转染的CHO细胞上清液中表达的sTNFRⅠ量超过1 000 ng/L,显著高于对照组(109.60±5.32) ng/L,P<0.001,表达的sTNFRⅠ能中和TNF-α对L929的细胞毒作用.结论 PcDNA3.1-sTNFRⅠ重组质粒在体外导入哺乳动物细胞后能够表达具有相应生物学活性的目的产物.
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