An Automated Induction Microfluidics System for Synthetic Biology,ACS Synthetic Biology

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An Automated Induction Microfluidics System for Synthetic Biology
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2018-03-08 00:00:00 , DOI: 10.1021/acssynbio.8b00025
Mathieu C. Husser _{1,

2} , Philippe Q. N. Vo ₃ , Hugo Sinha _{2,

3} , Fatemeh Ahmadi _{2,

3} , Steve C. C. Shih _{1,

2,

3}

Affiliation

The expression of a recombinant gene in a host organism through induction can be an extensively manual and labor-intensive procedure. Several methods have been developed to simplify the protocol, but none has fully replaced the traditional IPTG-based induction. To simplify this process, we describe the development of an autoinduction platform based on digital microfluidics. This system consists of a 600 nm LED and a light sensor to enable the real-time monitoring of the optical density (OD) samples coordinated with the semicontinuous mixing of a bacterial culture. A hand-held device was designed as a microbioreactor to culture cells and to measure the OD of the bacterial culture. In addition, it serves as a platform for the analysis of regulated protein expression in E. coli without the requirement of standardized well-plates or pipetting-based platforms. Here, we report for the first time, a system that offers great convenience without the user to physically monitor the culture or to manually add inducer at specific times. We characterized our system by looking at several parameters (electrode designs, gap height, and growth rates) required for an autoinducible system. As a first step, we carried out an automated induction optimization assay using a RFP reporter gene to identify conditions suitable for our system. Next, we used our system to identify active thermophilic β-glucosidase enzymes that may be suitable candidates for biomass hydrolysis. Overall, we believe that this platform may be useful for synthetic biology applications that require regulating and analyzing expression of heterologous genes for strain optimization.

中文翻译：

用于合成生物学的自动感应微流控系统

通过诱导在宿主生物中重组基因的表达可以是广泛的人工和劳动密集的过程。已经开发了几种方法来简化协议，但是没有一种方法可以完全替代传统的基于IPTG的归纳方法。为了简化此过程，我们描述了基于数字微流控技术的自动感应平台的开发。该系统由一个600 nm的LED和一个光传感器组成，能够实时监测与细菌培养物的半连续混合相协调的光密度（OD）样品。手持设备被设计为微生物反应器，用于培养细胞并测量细菌培养物的OD。此外，它还可以作为分析大肠杆菌中蛋白质表达调控的平台无需标准化的孔板或基于移液的平台。在此，我们首次报告了一种系统，该系统提供了极大的便利，而无需用户在特定时间进行物理监测培养或手动添加诱导剂。我们通过查看自动感应系统所需的几个参数（电极设计，间隙高度和生长速率）对系统进行了表征。第一步，我们使用RFP报告基因进行了自动化诱导优化分析，以鉴定适合我们系统的条件。接下来，我们使用我们的系统来识别可能适合生物质水解的活性嗜热性β-葡萄糖苷酶。全面的，

更新日期：2018-03-08

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