CAJ | 학술논문

目的通过原核表达系统高效制备人乳头瘤病毒（HPV）16晚期蛋白 L1病毒样颗粒（VLPs）。 　　方法构建 HPV16L1基因序列优化前后的 PET30a-HPV16L1重组质粒，转化大肠杆菌 Rosetta(DE3)；用 IPTG诱导目的基因表达，两步层析方法纯化 HPV16L1蛋白；电镜下观察纯化产物形成 VLPs 的情况。 　　结果成功构建大肠杆菌工程菌，高效可溶表达（目的蛋白约占总蛋白的38%）并纯化 HPV16L1蛋白，纯度达95%以上，电镜下观察，发现纯化后的目的蛋白为直径50 nm 左右，形态与天然病毒颗粒高度相似。 　　结论在大肠杆菌原核系统中高效、简易地制备了 HPV16L1 VLPs，为诊断试剂和疫苗的研制奠定基础。
목적통과원핵표체계통고효제비인유두류병독（HPV）16만기단백 L1병독양과립（VLPs）。 　　방법구건 HPV16L1기인서렬우화전후적 PET30a-HPV16L1중조질립，전화대장간균 Rosetta(DE3)；용 IPTG유도목적기인표체，량보층석방법순화 HPV16L1단백；전경하관찰순화산물형성 VLPs 적정황。 　　결과성공구건대장간균공정균，고효가용표체（목적단백약점총단백적38%）병순화 HPV16L1단백，순도체95%이상，전경하관찰，발현순화후적목적단백위직경50 nm 좌우，형태여천연병독과립고도상사。 　　결론재대장간균원핵계통중고효、간역지제비료 HPV16L1 VLPs，위진단시제화역묘적연제전정기출。
Objective To efficiently prepare HPV16L1 virus-like particles (VLPs) in Escherichia coli expression system. Methods Recombinant plasmids PET30a-HPV16L1 with condon usage optimization and non-optimization were constructed, and then transformed into E.coli Rosetta (DE3). IPTG was used to induce the recombinant E.coli Rosetta (DE3). By two step chromatography, the protein was purified. Electron microscopy was used to observe the structure of VLPs after purification. Results The recombinant engineered bacteria were successfully constructed. The solvable HPV16L1 protein (about 38% of the total protein) were efficiently expressed and purified, which reached the purity of higher than 95%. Self-assembling of VLPs for the purified L1 protein with the diameter of 50 nm was observed, showing high similarity with natural VLPs. Conclusion HPV16L1 VLPs was efficiently and conveniently prepared in Escherichia coli expression system, which will lay a foundation for further study of diagnostic reagent and vaccine.