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目的:构建人白细胞介素-24(hIL-24)基因的表达载体并在大肠杆菌中表达。方法:用PCR从质粒TRAP-IL-24中扩增hIL-24cDNA片段,并将该片段插入pGEX-KG原核表达载体中,实现插入基因的融合表达。用SDS-PAGE和West-ernblot对表达产物进行鉴定。采用MTT法检测GST-IL-24融合蛋白的生物学活性。结果:酶切结果证实,成功地构建了pGEX-KG IL24原核表达载体,并在大肠杆菌中获得稳定的表达,表达产物的相对分子质量(Mr)同预期值相一致。GST-IL24融合蛋白能够抑制THP1细胞的生长。结论:成功地构建了重组表达载体pGEX-KG-IL24,并在E.coliBL21中表达了具有生物学活性的GST-IL-24融合蛋白,为下一步研究人IL24的功能奠定了基础。
Objective: To construct an expression vector for human interleukin-24 (hIL-24) gene and express in E. coli. Methods: hIL-24 cDNA fragment was amplified from plasmid TRAP-IL-24 by PCR. The fragment was inserted into pGEX-KG prokaryotic expression vector to express the fusion gene. The expression products were identified by SDS-PAGE and West-ernblot. The biological activity of GST-IL-24 fusion protein was detected by MTT assay. Results: The results of enzyme digestion confirmed that prokaryotic expression vector pGEX-KG IL24 was successfully constructed and expressed stably in E. coli. The relative molecular mass (Mr) of the expressed product was consistent with the expected value. GST-IL24 fusion protein can inhibit the growth of THP1 cells. CONCLUSION: The recombinant expression vector pGEX-KG-IL24 was successfully constructed and the biologically active GST-IL-24 fusion protein was expressed in E. coli BL21, which laid the foundation for the further study on the function of human IL24.