一组焦亡执行者融合蛋白和基于荧光素酶片段互补分析技术的焦亡活性检测方法

• Format: Patent
• Language: Chinese
• Published: 25.03.2022
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS OR TEST PAPERS THEREFOR; COMPOSITIONS THEREOF; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; CULTURE MEDIA; ENZYMOLOGY; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PROCESSES OF PREPARING SUCH COMPOSITIONS; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

本发明提供了一组焦亡执行者融合蛋白和基于荧光素酶片段互补分析技术的焦亡活性检测方法，属于细胞生物学技术领域。焦亡执行者融合蛋白，包括hGlucN-GSDME融合蛋白和hGlucC-GSDME融合蛋白。一组焦亡活性报告质粒，包括分别含上述两融合蛋白的编码序列的真核表达载体。所述融合蛋白或所述报告质粒在GSDME介导的细胞焦亡活性检测、制备检测GSDME介导的细胞焦亡活性的试剂或试剂盒或GSDME介导焦亡诱导剂或抑制剂筛选中的应用。本发明首次将荧光素酶片段互补分析技术应用于细胞焦亡活性检测中，对GSDME激活水解产物N-GSDME的聚集进行检测，为研究细胞焦亡提供了新的技术方法。 The invention provides a group of pyroptosis executor fusion proteins and a pyroptosis activity detection method based on a luciferase fragment complementation assay technology, and belongs to the technical field of cell biology. The pyroptosis executor fusion proteins comprise an hGlucN-GSDME fusion protein and an hGlucC-GSDME fusion protein. A group of pyroptosis activity reporter plasmids comprise eukaryotic expression vectors respectively containing the coding sequences of the two fusion proteins. The fusion proteins or the reporter plasmids are applied to GSDME-mediated pyroptosis activity detection, preparation of a reagent or kit for detecting GSDME-mediated pyroptosis activity or screening of a GSDME-mediated pyrop