Research

Synthesizing non-natural parts from natural genomic template

Pawan K Dhar¹ , Chaw Su Thwin¹ , Kyaw Tun¹ , Yuko Tsumoto¹ , Sebastian Maurer-Stroh² , Frank Eisenhaber² and Uttam Surana³

¹  Synthetic Biology Lab, RIKEN Advanced Sciences Institute, Yokohama, 230-0045, Japan

²  Biomolecular Function Discovery Division, Bioinformatics Institute, Agency for Science, Technology and Research (A*STAR), 30 Biopolis Street, 138673, Singapore

³  Cell Cycle control Lab, Institute of Molecular and Cellular Biology, 61 Biopolis Drive, 138673, Singapore

author email corresponding author email

Journal of Biological Engineering 2009, 3:2doi:10.1186/1754-1611-3-2

Published:	3 February 2009

Abstract

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.

Conclusion

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.