An Automated Induction Microfluidics System for Synthetic Biology

合成生物学 诱导剂 微流控 计算生物学 异源的 移液管 紫胶操纵子 计算机科学 系统生物学 生物 基因表达 基因 纳米技术 生化工程 化学 遗传学 工程类 材料科学 物理化学
作者
Mathieu C. Husser,Philippe Q. N. Vo,Hugo Sinha,Fatemeh Ahmadi,Steve C. C. Shih
出处
期刊:ACS Synthetic Biology [American Chemical Society]
卷期号:7 (3): 933-944 被引量:28
标识
DOI:10.1021/acssynbio.8b00025
摘要

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.

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