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Novel T7-like expression systems used for Halomonas.

Metab. Eng.. 2017-01; 
ZhaoHan, ZhangHaoqian M, ChenXiangbin, LiTeng, WuQiong, OuyangQi, ChenGuo-Q
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Gene Synthesis … T7-family RNAPs and promoters (P T7-like ) were de novo synthesized at GenScript (Nanjing, China), cloned into RNAP-module plasmid (p15A origin, Amp + ) and promoter-module plasmid (pSC101 origin, Cm + ) respectively, via the Golden Gate Assembly method (see Table … Get A Quote

摘要

To engineer non-model organisms, suitable genetic parts must be available. However, biological parts are often host strain sensitive. It is therefore necessary to develop genetic parts that are functional regardless of host strains. Here we report several novel phage-derived expression systems used for transcriptional control in non-model bacteria. Novel T7-like RNA polymerase-promoter pairs were obtained by mining phage genomes, followed by in vivo characterization in non-model strains Halomonas spp TD01 and Pseudomonas entomophila. Three expression systems, namely, MmP1, VP4, and K1F, were developed displaying orthogonality (crosstalk<0.7%), tight regulation (3085-fold induction), and high... More

关键词

Expression system,Morphological engineering,Non-model microorganism,PHB,Part mining,Synthetic bio