초록

본 연구개발은 기존 미생물 단백질 발현 시스템을 분석하고 재구성하는 시스템합성 융복합 연구를 통해, 의약 산업 및 농수축산업에의 활용을 아우르는 고품질 단백질 생산용 플랫폼 균주를 개발하였음. 이를 위해, 1) 무항생제 기반 인공 플라스미드를 위한 신규 제어인자 발굴 및 발현량 프로그래밍을 통해 복제수 조절 시스템 및 2) 자연계의 전사-번역 동조 시스템 재구성을 통해 비손상 주형 특이적 번역 개시 제어 시스템을 개발함으로써 최종 목표를 달성하였음.

(출처 : 요약서 3p)

Abstract ▼

□ Purpose&Contents
This R&D has developed a platform strain for the production of high-quality proteins used in the pharmaceutical, agricultural, and livestock industries through systems and synthetic biologythat analyze and reconstruct existing microbial protein expression systems. To this end,

□ Purpose&Contents
This R&D has developed a platform strain for the production of high-quality proteins used in the pharmaceutical, agricultural, and livestock industries through systems and synthetic biologythat analyze and reconstruct existing microbial protein expression systems. To this end, 1) a system to control the number of copies of plasmid through discovery and expression level programming of new selection factors for artificial antibiotics-free plasmids and 2) reconfiguration of natural bacterial expression system to decoupling transcription
-translation system were developed.

□ Results
A plasmid copy number dynamic control system based on the antibiotics-free artificial plasmid
-It was confirmed that both artifitial antibiotics-free plasmids having CloDF (medium copy number) and pUC origin (high copy number), respectivly, were stably maintained in cells without antibiotics when the infA gene was used as a selection marker. In addition, the plasmid capable of dynamic regulation of plasmid copy number was constructed based on an antibiotic-free plasmid system using an inducible promoter to control selection marker expression.
Intact template-specific translation initiation control system
-Synthetic expression cassettes that control the translation initiation to start translation only with intact mRNA has been developed. It was verified that the quality control of the expressed protein is possible with the developed expression system by comparing the amount of N terminus with C terminus of the long fusion protein. The developed ProQC system expressed more high-quality protein compared to the natural system (1.6 times the protein of full length).

□ Expected Contribution
This study made it possible to 1) construction of an artificial plasmid system capable to be designed to have multiple copies even with a single origin of replication 2) commercialization of plasmids incorporating high-quality protein expression cassettes with decoupled transcription –translation system.
-The developed high-quality protein production platform strain can be introduced not only in the process of using the protein itself as a final product, but also in metabolic pathways that increase productivity using fusion protein technology. In the case of a fusion protein, the length of the protein is usually long, so producing a high-quality protein using the ProQC system willbe able to increases production efficiency.

(출처 : SUMMARY 6p)

목차 Contents

• 표지 ... 1
• 제출문 ... 2
• 보고서 요약서 ... 3
• 국 문 요 약 문 ... 4
• Summary ... 6
• 목차 ... 7
• 제 1 장 연구개발과제의 개요 ... 8
• 제 1 절 연구개발 목적 ... 8
• 제 2 절 연구개발의 필요성 ... 8
• 제 3 절 연구 개발 범위 ... 9
• 제 2 장 연구수행 내용 및 결과 ... 10
• 제 1 절 이론적, 실험적 접근방법 ... 10
• 제 2 절 연구 수행 내용 및 결과 ... 12
• 제 3 장 목표달성도 및 관련분야 기여도 ... 21
• 제 1 절 목표대비 달성도 ... 21
• 제 2 절 정량적 성과(논문게재, 특허출원, 기타)를 기술 ... 21
• 제 4 장 연구결과의 활용계획 ... 22
• 제 5 장 연구개발 결과의 보안 등급 ... 23
• 제 6 장 연구시설·장비종합정보시스템에 등록한 연구시설·장비 현황 ... 23
• 제 7 장 연구개발과제의 대표적 연구실적 ... 24
• 제 8 장 기타사항 ... 24
• 제 9 장 참고문헌 ... 25
• 끝페이지 ... 26