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Rutgers scientists have recently devised a novel expression system to produce mRNA transcripts without the detrimental stem-loop structure near the translation initiation site.
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Summary: The synthesis of recombinant proteins using prokaryotic expression systems has been instrumental in producing valuable reagents for biomedical research and commercial products. However, the majority of in vivo expression systems have inherent limitations in most applications--including low yields, poor quality of the expressed proteins and the inability of common expression vectors such as E.coli to translate "rare codons" from other organisms. Rutgers scientists have recently devised a novel expression system to produce mRNA transcripts without the detrimental stem-loop structure near the translation initiation site and optimal codon usage. This technology enables both the design and customization of the desired expression vector within the host cells using a technique called "Transcript-Optimized Expression Enhancement Technology (TOEET)". In this expression platform, DNA sequences are customized to produce mRNA transcripts with minimal secondary structure in the nucleotide region around the Ribosomal Binding Site (RBS), along with the maximal codon usage near the N-terminal region. This leads to enhanced translation initiation and protein expression capacities. Further, the sequences can be engineered to yield polypeptides featuring enhanced solubility and purification capacities. This technology addresses several limitations in the production of extremely valuable recombinant proteins and better enables biotechnologists to design more effective processes for producing proteins at industrial scales. In summary, this comprehensive protein expression system delivers superior-quality and high quantity of recombinant proteins from the genes of interest, which can be used for several biomedical research and therapeutic applications. Advantages: TOEET technology eliminates the need for optimizing target gene sequence, thereby avoiding gene-specific synthesis or modification. This technology enables synthesis of large amounts of proteins without the need for the high cost total gene synthesis.