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目的构建重组质粒pcDNA3.1/PGRP,肌肉注射免疫Balb/c小鼠,探讨PGRP的基因免疫效果。方法胶回收纯化的PGRP基因片段及pcDNA3.1质粒经HindⅢ、BamHⅠ双酶切,T4DNA Ligase连接构建成重组质粒pcDNA3.1/PGRP,用PCR方法对其进行鉴定,质粒抽提后进行序列分析;于小鼠后腿双侧肌注免疫,检测各项免疫指标。结果构建的重组质粒经PCR鉴定和测序证明PGRP插入方向正确,免疫检测结果显示重组质粒激活了PGRP特异的CTL和Th1细胞,同时PGRP特异性抗体效价小幅升高。结论构建的重组质粒pcDNA3.1/PGRP具有诱导特异性体液免疫应答和细胞免疫应答的能力,可作为潜在的小细胞肺癌新型疫苗,有进一步研究的意义。
Objective To construct recombinant plasmid pcDNA3.1 / PGRP and immunize Balb / c mice intramuscularly to investigate the gene immune effect of PGRP. Methods Plasmid pGRP gene fragment was recovered by PCR and pcDNA3.1 plasmid was digested with Hind Ⅲ and BamH Ⅰ. The recombinant plasmid pcDNA3.1 / PGRP was constructed by T4 DNA Ligase and identified by PCR. The plasmid was extracted and sequenced. Mice immunized with bilateral intramuscular injection of hind legs, the detection of various immune indicators. Results The recombinant plasmids were verified by PCR and sequencing. The results of immunoassay showed that PGRP-specific CTL and Th1 cells were activated by recombinant plasmids, and the titer of PGRP-specific antibody increased slightly. Conclusion The constructed recombinant plasmid pcDNA3.1 / PGRP has the ability of inducing specific humoral and cellular immune responses and may serve as a potential new vaccine for small cell lung cancer. It is of great significance to further study.