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A combined cell-free transcription-translation system from Saccharomyces cerevisiae for rapid and robust protein synthesis.

Biotechnol J.. 2014-02; 
R Gan, MC Jewett. Department of Chemical and Biological Engineering, Northwestern University, 2145 Sheridan Road, Evanston, IL 60208, USA.
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摘要

Cell-free protein synthesis (CFPS) provides a valuable platform for understanding, using, and expanding the capabilities of the translation apparatus. For example, high-throughput CFPS is helping to address the increasing discrepancy between genome sequence data and their translation products. Here, we report the development of a combined cell-free transcription and translation system from Saccharomyces cerevisiae that is suitable for such efforts. First, we show the ability to enable translation initiation in a cap-independent manner. The performance of various genetic elements were assessed, including 5"-UTR, 3"-UTR, and length of poly(A) tail. A specific vector harboring the 5"-UTR fragment of... More

关键词

yeast; combined transcription-translation; cell-free protein synthesis; in vitro