Synthesizing non-natural parts from natural genomic template.
BACKGROUND
The current knowledge of genes and proteins comes from 'naturally designed' coding and non-coding regions. It would be interesting to move beyond natural boundaries and make user-defined parts. To explore this possibility we made six non-natural proteins in E. coli. We also studied their potential tertiary structure and phenotypic outcomes.


RESULTS
The chosen intergenic sequences were amplified and expressed using pBAD 202/D-TOPO vector. All six proteins showed significantly low similarity to the known proteins in the NCBI protein database. The protein expression was confirmed through Western blot. The endogenous expression of one of the proteins resulted in the cell growth inhibition. The growth inhibition was completely rescued by culturing cells in the inducer-free medium. Computational structure prediction suggests globular tertiary structure for two of the six non-natural proteins synthesized.


CONCLUSIONS
To our best knowledge, this is the first study that demonstrates artificial synthesis of non-natural proteins from existing genomic template, their potential tertiary structure and phenotypic outcome. The work presented in this paper opens up a new avenue of investigating fundamental biology. Our approach can also be used to synthesize large numbers of non-natural RNA and protein parts for useful applications.
Dhar
Pawan K
PK
Synthetic Biology Lab, RIKEN Advanced Sciences Institute, Yokohama, 230-0045, Japan. pkdhar@riken.jp
Thwin
Chaw Su
CS
Tun
Kyaw
K
Tsumoto
Yuko
Y
Maurer-Stroh
Sebastian
S
Eisenhaber
Frank
F
Surana
Uttam
U