Redesign or modification of the cellular physiology requires a quantitatively well-controlled expression system known as the “tunable expression.” Although the modification of promoters demonstrates the great impact on the translation efficiency, it is difficult to detect the proper variants required for tunable expression. The 5′-untranslated region (UTR), however, can be an important target for tunable expressions because the ribosome binding affinity is directly modulated by the sequence variants of the Shine-Dalgarno (SD) sequence and the AU-rich sequence, which are the ribosome binding sites and a SD-sequence-independent translation enhancer, respectively. Additionally, downstream region (DR) of translation initiation codon also affect on the expression level depending on its secondary structure. This study developed a simple method to obtain numerous 5′UTR variants and analyze their translation efficiency based on the PCR-based site-directed mutagenesis and the expressional PCR using coupled in vitro transcription/ translation system derived from E. coli and eGFP gene as a template. SD sequence variants and AU-rich sequence variants, which have a wide range of relative expression levels ranging from 0.1 to 2.0, were obtained. Furthermore, silent mutations around DR showed a significant difference in the expression level more than 10 times. We found that the translation efficiency was affected by the ribosome binding affinity and its accessibility that is dependent on the secondary structure around the 5′UTR and DR. The results in this study illustrates the potentials fine control of expression level based on the modifications in 5’ UTR and DR structures.
저자
Gyoo Yeol Jung [ Department of Chemical Engineering, School of Interdisciplinary Bioscience and Bioengineering, POSTECH, Pohang, Korea ]
Sang Woo Seo [ Department of Chemical Engineering ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
이 법인은 생물 공학의 발전과 보급에 이바지하고, 회원 상호 간의 연구 협력과 친목을 도모함을 목적으로 한다
1. 생물공학 분야의 발전을 위한 연구 협력
2. 생물공학의 실용화를 촉진시키기 위한 산학 협동
3. 학술연구 발표회, 강연회, 연수회 등 학술활동의 개최
4. 국,영문 학술지,소식지,학술회의 Proceedings 및 학술도서의 발간
5. 생물공학 발전을 위한 정책 건의
6. 기타 국제 교류 등 본 학회의 목적 달성을 위한 제반 활동