期刊
ACS SYNTHETIC BIOLOGY
卷 6, 期 4, 页码 638-647出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssynbio.6b00250
关键词
cell-free synthetic biology; transcription-translation; IVTT; PURE system; artificial cell
资金
- Armenise-Harvard foundation
- CIBIO
- Autonomous Province of Trento, Grandi Progetti, project Characterizing and improving brain mechanisms of attention-ATTEND
Although RNA synthesis can be reliably controlled with different T7 transcriptional promoters during cell-free gene expression with the PURE system, protein synthesis remains largely unaffected. To better control protein levels, we investigated a series of ribosome binding sites (RBSs). Although RBS strength did strongly affect protein synthesis, the RBS sequence could explain less than half of the variability of the data. Protein expression was found to depend on other factors besides the strength of the RBS, including the GC content of the coding sequence. The complexity of protein synthesis in comparison to RNA synthesis was observed by the higher degree of variability associated with protein expression. This variability was also observed in an E. coli cell extract-based system. However, the coefficient of variation was larger with E. coli RNA polymerase than with T7 RNA polymerase, consistent with the increased complexity of E. coli RNA polymerase.
作者
我是这篇论文的作者
点击您的名字以认领此论文并将其添加到您的个人资料中。
推荐
暂无数据