Integrating T7 RNA Polymerase and Its Cognate Transcriptional Units for a Host-Independent and Stable Expression System in Single Plasmid
Integrating T7 RNA Polymerase and Its Cognate Transcriptional Units for a Host-Independent and Stable Expression System in Single Plasmid
复制标题
整合 T7 RNA 聚合酶及其同源转录单位，在单质粒中构建宿主独立且稳定的表达系统
DOI：
10.1021/acssynbio.8b00055
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发表时间：
2018-05-01
影响因子：
4.7
通讯作者：
Li, Qiang
中科院分区：
文献类型：
--
作者：
Liang, Xiao;Li, Chenmeng;Li, Qiang
Metabolic engineering and synthetic biology usually require universal expression systems for stable and efficient gene expression in various organisms. In this study, a host-independent and stable T7 expression system had been developed by integrating T7 RNA polymerase and its cognate transcriptional units in single plasmid. The expression of T7 RNA polymerase was restricted below its lethal threshold using a T7 RNA polymerase antisense gene cassette, which allowed long periods of cultivation and protein production. In addition, by designing ribosome binding sites, we further tuned the expression capacity of this novel T7 system within a wide range. This host-independent expression system efficiently expressed genes in five different Gram-negative strains and one Gram-positive strain and was also shown to be applicable in a real industrial D-p-hydroxyphenylglycine production system.
