AI Article Synopsis
- Researchers have developed a method called "TED," which enhances protein production in E. coli by using a gene sequence from the slime mold Dictyostelium discoideum.
- By placing this gene sequence near the ribosome binding site of a target gene, protein expression levels can be significantly increased, specifically the mlcR gene showed the most promise.
- Testing showed that integrating TED with a T7 expression system resulted in higher protein yields compared to traditional methods, offering a new tool to improve biotechnological applications.
Article Abstract
Methods for heterologous protein production in Escherichia coli have revolutionized biotechnology and the bioindustry. It is ultimately important to increase the amount of protein product from bacteria. To this end, a variety of tools, such as effective promoters, have been developed. Here, we present a versatile molecular tool based on a phenomenon termed "translation enhancement by a Dictyostelium gene sequence" ("TED") in E. coli. We found that protein expression was increased when a gene sequence of Dictyostelium discoideum was placed upstream of the Shine-Dalgarno sequence located between the promoter and the initiation codon of a target gene. The most effective sequence among the genes examined was mlcR, which encodes the myosin regulatory light chain, a subunit of myosin II. Serial deletion analysis revealed that at least 10 bases of the 3' end of the mlcR gene enhanced the production of green fluorescent protein in cells. We applied this tool to a T7 expression system and found that the expression level of the proteins tested was increased when compared with the conventional method. Thus, current protein production systems can be improved by combination with TED.
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| http://dx.doi.org/10.1007/s00253-019-09746-7 | DOI Listing |
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