CAJ | 학술논문

家蚕核型多角体病毒在感染晚期大量表达产生多角体蛋白,并形成结构致密的超大分子蛋白质晶体,以有效保护内部包埋的病毒粒子,保证子代病毒的传播.借鉴多角体包埋病毒粒子的原理,观察到多角体也能将外源蛋白质固定入多角体内部.利用能形成多角体的BmNPV PolH+Bac-to-Bac表达系统构建的含有绿色荧光蛋白(EGFP)的重组病毒能在表达、产生正常多角体的同时也表达EGFP蛋白.共聚焦分析表明，该重组病毒感染后形成的多角体能够发射绿色荧光.被包埋的EGFP蛋白放置1个月后仍能检测到绿色荧光,干燥处理30 min后仍能检测到绿色荧光.由此说明将多角体蛋白与外源基因编码蛋白同时表达,多角体中会包埋外源蛋白,且能较长时间保存外源蛋白的生物活性.这一现象为解决蛋白质长期保存提供了新思路,对于开发含有毒素蛋白的新型生物杀虫剂也有很好的参考意义.
가잠핵형다각체병독재감염만기대량표체산생다각체단백,병형성결구치밀적초대분자단백질정체,이유효보호내부포매적병독입자,보증자대병독적전파.차감다각체포매병독입자적원리,관찰도다각체야능장외원단백질고정입다각체내부.이용능형성다각체적BmNPV PolH+Bac-to-Bac표체계통구건적함유록색형광단백(EGFP)적중조병독능재표체、산생정상다각체적동시야표체EGFP단백.공취초분석표명，해중조병독감염후형성적다각체능구발사록색형광.피포매적EGFP단백방치1개월후잉능검측도록색형광,간조처리30 min후잉능검측도록색형광.유차설명장다각체단백여외원기인편마단백동시표체,다각체중회포매외원단백,차능교장시간보존외원단백적생물활성.저일현상위해결단백질장기보존제공료신사로,대우개발함유독소단백적신형생물살충제야유흔호적삼고의의.
In late phase of BmNPV infection,the virus produces large amount of polyhedrin and forms polyhedra, which contain compact protein crystals. Polyhedron is highly stable, enabling the incorporated virions to persist in unfavorable environment and to have seasonal infections to host larvae. In this study,we investigated whether the foreign protein co-expressed with polyhedrin in recombinant BmNPV could be incorporated into polyhedra like virions. Recombinant baculovirus expressing polyhedrin and EGFP simultaneously was constructed. The results from electron microscopy observation showed that the polyhedra formed from the recombinant baculovirus could incorporate virion as the wild-type BmNPV. Green fluorescence was detected on the surface of purified polyhedra under confocal microscopy. Dissolved BmNPV-EGFP polyhedra emitted green fluorescence after desiccation treatment for 30 min. EGFP bioactivity could also be detected in polyhedra under room temperature after one month. The observation from this experiment showed that the foreign protein, EGFP, could be embedded into polyhedra when co-expressed with polyhedrin and its bioactivity can persist for longer time. It provided a new inspiration for efficient preservation of useful protein and for development of new biopesticides containing toxin proteins.