大肠癌新型CEA疫苗重组杆状病毒的表达与鉴定

Construction and identification of recombinant baculovirus of the novel CEA vaccine

目的利用Bac-to-Bac杆状病毒表达系统,构建新型CEA融合表位疫苗(以HPV16L1为蛋白载体)的重组杆状病毒。方法构建含有HPV16L1片段和CEA-PADRE合成片段的表达载体pFastBac1-V,转化DH10Bac感受态菌。利用其含有的细菌Tn7转座系统,将该基因重组至杆状病毒穿梭质粒bacm id中,转染昆虫细胞sf9,获得新型CEA疫苗的重组杆状病毒。结果限制性内切酶酶切和DNA序列分析表明目的片断正确克隆到杆状病毒转移载体pFastBac1中;重组杆状病毒感染昆虫细胞后,PCR检测可以扩增出相应大小的片段。结论利用杆状病毒表达系统,成功构建了CEA融合表位疫苗的重组杆状病毒,为进一步的研究奠定了基础。

Objective To design a novel efficient CEA vaccine of fission epitopes with the vehicle of HPV16L1, and construct the recombinant baculovirus in Bac-to-Bac baculovirus expression system. Methods The recombinant baculovirus expression vector pFastBacl-V was constructed. HPV16L1 and CEA-PADRE synthesis primers were inserted into bacmid in E. coli DH10Bac with the help of Tn7 transposition system. Then the recombinant baculovirus was got by infecting insect cell sf9. Results HPV16L1 and CEA-PADRE synthesis primers that were cloned to the baculovirus transfer vector pFastBacl were assessed by restriction analysis and direct sequencing. The recombinant baculovirus of the novel vaccine had been constructed. After infected sf9 ,the correct fragment was amplified by PCR. Conclusion The baculovirus of CEA vaccine of fussion epitopes with the vehicle of HPV16L1 was constructed.