Tuning the expression level of recombinant proteins by modulating mRNA stability in a cell-free protein synthesis system.
In this work, it was discovered that the stability of mRNA in a cell-free extract could be controlled by using engineered T7 terminator sequences. Specifically, it was found that mRNA stability gradually decreased as the length of the stem structure of the T7 terminator was reduced sequentially. As a result of the controlled abundance of mRNA species, it was possible to manipulate the relative expression level of target proteins by employing the T7 terminator of adjusted stem lengths.
Ahn
Jin-Ho
JH
Institute of Molecular Biology and Genetics, Seoul National University, Seoul, Korea.
Kang
Taek-Jin
TJ
Kim
Dong-Myung
DM
D017123:Bacteriophage T7 / Q000235:genetics; D002474:Cell-Free System; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000096:biosynthesis; D020871:RNA Stability; D012333:RNA, Messenger / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013728:Terminator Regions, Genetic; D014764:Viral Proteins / Q000235:genetics
D012333:RNA, Messenger; D011994:Recombinant Proteins; D014764:Viral Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins