The baculovirus expression system (BES) utilize the p10 or polyhedrin promoter, a very late promoter that exhibits strong transcriptional activity primarily at the end of viral infection, to produce useful recombinant proteins. The burst sequence of the very late promoter is essential for strong transcription, and VLF-1 is a transcription factor that binds specifically to the burst sequence, and it has been reported that it can regulate the amount and timing of expression of protein by the very late promoter. Recently, a VLF-1 constitutively expressing cell line was constructed to increase the production of the target protein, but the effect was minimal. In this study, to find the optimal VLF-1 expression conditions to increase target protein production efficiency, we controlled the expression of VLF-1 through various promoters and evaluated the target protein expression efficiency by the p10 promoter accordingly.

Hand, foot, and mouth disease (HFMD) is a highly contagious disease with no specific treatment. Since it is common in immunocompromised children under the age of 5, there is a need to develop a safe vaccine. Virus-like particles (VLPs) are similar structures to viruses with the lack of genetic material which makes them impossible to replicate and infect, and therefore have a high level of biological safety and are considered to have high value as vaccines. In this study, the insect virus expression system that is widely used for vaccine and drug production due to its high post-translational modification efficiency, was used to produce VLPs for Coxsackievirus type A6 and A10, which are recently reported to be the main causes of HFMD. For this purpose, the selection of promoters that can control the timing and intensity of expression of 3CD protein, which is essential for VLPs assembly but has been reported to be cytotoxic, was conducted to construct an optimal expression form for HFMD-VLP.

표면발현(surface-display system)은 세포 또는 바이러스 표면에 목적 단백질을 고정하여 발현시킴으로써 목적 단백질에 대하여 독립적인 공간 구조 및 생물학적 활성을 부여하는 단백질 공학 기술이다. 또한 이를 이용하여 높은 중화항체 유도 및 대량생산이 가능한 삼량체의 형태로 항원 단백질의 발현 또한 가능하다. BES(baculovirus expression system)에서의 표면발현 기술은 번역 후 수정과정 및 복잡한 구조의 다양한 단백질의 발현이 가능하기 에 다른 숙주 기반 시스템보다 효율적이라고 보고되고 있다. 그러나 목적 단백질 외의 다른 표면 단백질과 발현 공간에서의 경쟁으로 목적 단백질의 낮은 생산량이 큰 문제점으로 지적되고 있다. 따라서, 이러한 BES에서 표면 발현의 생산 효율을 증대시키기 위하여, 동일한 표면 공간에 대한 단백질 간의 발현 경쟁에 대해 실험적으로 확인 후, 그를 해결하기 위하여 표면발현에 최적인 목적 단백질 발현을 위한 프로모터 선발 실험을 수행하였다. 이를 통해 BES에서 표면발현에 의한 목적 단백질의 생산 효율을 증대시킬 수 있음을 확인하였다.