Protocol to develop a synthetic biology toolkit for the non-model bacterium R. palustris.
Protocol to develop a synthetic biology toolkit for the non-model bacterium R. palustris.
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DOI：
10.1016/j.xpro.2023.102158
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发表时间：
2023-04-26
期刊：
影响因子：
--
通讯作者：
Saha, Rajib
中科院分区：
文献类型：
--
作者：
Kathol, Mark;Immethun, Cheryl;Saha, Rajib
许多生物学工具被开发出来用于模式生物，然而，这些工具在非模式生物中并不有效。在这里，我们提出了为沼泽红假单胞菌CGA009开发合成生物学工具包的方案，沼泽红假单胞菌是一种具有独特代谢特性的非模式细菌。我们描述了在非模式细菌中引入和表征生物装置的步骤，例如利用荧光标记和RT-qPCR。该方案也可适用于其他非模式生物。有关本议定书的使用和执行的完整细节，请参阅Immetun等人。适用于表征非模式生物中生物装置的协议沼泽假单胞菌中异源基因表达的优化合成生物学工具箱将异源基因整合到沼泽假单胞菌的内源质粒出版商的说明：进行任何实验方案都需要遵守当地实验室安全和伦理的机构指南。许多生物学工具被开发出来用于模式生物，但这些工具在非模式生物中并不有效。在这里，我们提出了为沼泽红假单胞菌CGA009开发合成生物学工具包的方案，沼泽红假单胞菌是一种具有独特代谢特性的非模式细菌。我们描述了在非模式细菌中引入和表征生物装置的步骤，例如利用荧光标记和RT-qPCR。该方案也可适用于其他非模式生物。
Numerous biology tools are developed to work for model organisms, which, however, do not work effectively in non-model organisms. Here, we present a protocol for developing a synthetic biology toolkit for Rhodopseudomonas palustris CGA009, a non-model bacterium with unique metabolic properties. We describe steps for introducing and characterizing biological devices in non-model bacteria, such as the utilization of fluorescence markers and RT-qPCR. This protocol may also be applicable for other non-model organisms. For complete details on the use and execution of this protocol, please refer to Immethun et al.. Protocol applicable for characterizing biological devices in non-model organisms Optimized synthetic biology toolbox for heterologous gene expression in R. palustris Steps for incorporating heterologous genes into R. palustris’ endogenous plasmid Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Numerous biology tools are developed to work for model organisms, which however do not work effectively in non-model organisms. Here, we present a protocol for developing a synthetic biology toolkit for Rhodopseudomonas palustris CGA009, a non-model bacterium with unique metabolic properties. We describe steps for introducing and characterizing biological devices in non-model bacteria, such as the utilization of fluorescence markers and RT-qPCR. This protocol may also be applicable for other non-model organisms.
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