A Dual-Replicon Shuttle Vector System for Heterologous Gene Expression in a Broad Range of Gram-Positive and Gram-Negative Bacteria
A Dual-Replicon Shuttle Vector System for Heterologous Gene Expression in a Broad Range of Gram-Positive and Gram-Negative Bacteria
复制标题
用于在广泛的革兰氏阳性和革兰氏阴性细菌中表达异源基因的双复制子穿梭载体系统
DOI：
10.1007/s00284-018-1535-8
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发表时间：
2018-07
影响因子：
2.6
通讯作者：
Zeng Hui
中科院分区：
文献类型：
--
作者：
Hua Mingxi;Guo Jingjing;Li Min;Chen Chen;Zhang Yuanyuan;Song Chuan;Jiang Dong;Du Pengcheng;Zeng Hui
复制的起源(在滚动的圆圈复制质粒中的蛋黄素-theta复制质粒)在广泛的细菌中启动质粒复制。这两种质粒都在链球菌中发现，链球菌是一种由人类共生细菌和病原体组成的属，能够引起严重的社区获得性感染，包括脑膜炎、败血症和呼吸道疾病。鉴于链球菌在与其他共生微生物交换遗传元素方面的重要作用，我们对两种链球菌的基因型别和表型进行了研究。和其他共生物种在寄主定殖过程中可能发生变化。因此，需要一种改进的质粒系统来研究链球菌功能、复杂和多变的基因组。本研究构建了一个双复制子穿梭载体系统pDRE以实现异源基因的表达。该载体系统含有大肠杆菌的theta复制子。在革兰氏阳性菌中根据pMV158合成了滚环复制子的起源。通过检测插入基因在多个克隆位点的产物，评估了该载体系统在四种链球菌和另外三种革兰氏阳性菌：枯草杆菌、乳酸乳球菌和金黄色葡萄球菌中复制和表达异源基因的能力。结果表明，新构建的载体可以在多种革兰氏阳性和革兰氏阴性菌中同时复制和表达异源基因，为进一步的功能分析提供了一个潜在的有力的遗传工具。
Origin of replication (oriin theta-replicating plasmids ordsoin rolling circle replicating plasmids) initiates plasmid replication in a broad range of bacteria. These two kinds of plasmids were both identified inStreptococcus, a genus composed of both human commensal bacteria and pathogens with the ability to cause severe community-acquired infections, including meningitides, septicemia, and respiratory tract diseases. Given the important roles ofStreptococcusin the exchange of genetic elements with other symbiotic microbes, the genotypes and phenotypes of bothStreptococcusspp. and other symbiotic species could be changed during colonization of the host. Therefore, an improved plasmid system is required to study the functional, complicated, and changeable genomes ofStreptococcus. In this study, a dual-replicon shuttle vector system named pDRE was constructed to achieve heterologous gene expression. The vector system contained theta replicon forEscherichia coli. The origin of rolling circle replicon was synthesized according to pMV158 in Gram-positive bacteria. By measuring the products of inserted genes at multiple cloning sites, the ability of this vector system in the replication and expression of heterologous genes was assessed in fourStreptococcusand three other Gram-positive bacteria:Bacillus subtilis, Lactococcus lactis, andStaphylococcus aureus. The results showed that the newly constructed vector could simultaneously replicate and express heterologous genes in a broad range of Gram-positive and Gram-negative bacteria, thus providing a potentially powerful genetic tool for further functional analysis.
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影响因子：
4.9
作者：
Jensen, Anders;Valdorsson, Oskar;Kilian, Mogens
通讯作者：
Kilian, Mogens
DOI：
--
发表时间：
--
期刊：
--
影响因子：
--
作者：
G. Solar;Rafael Giraldo;M. Ruiz-Echevarría;M. Espinosa;R. Dı́az-Orejas
通讯作者：
G. Solar;Rafael Giraldo;M. Ruiz-Echevarría;M. Espinosa;R. Dı́az-Orejas
影响因子：
48
作者：
Gibson, Daniel G.;Young, Lei;Smith, Hamilton O.
通讯作者：
Smith, Hamilton O.
影响因子：
14.9
作者：
Bode M;Khor S;Ye H;Li MH;Ying JY
通讯作者：
Ying JY
影响因子：
11.3
作者：
Willems RJ;Hanage WP;Bessen DE;Feil EJ
通讯作者：
Feil EJ