Identification of a gene that makes a protein by using a cell free protein synthesis system

A living cell could be genetically modified to perform a function such as the production of a protein. However, these genetic modifications often conflict with normal cellular function and result in a mutation. Defects can be overcome through removing the bacterial membrane which leaves the lysate that is performing both transcription and translation. The cell free-protein synthesis is also known as in vitro protein synthesis and is the production of a protein without using a living cell. The gene is acting as instructions to make the protein. If we can isolate a gene and then apply a cell free protein synthesis system after synthesis the protein and run on gel-electrophoresis we can identify a gene on the basis of the protein. Gel electrophoresis is a laboratory technique used to ______ contrasting proteins according to molecular size and charge.

Microfluidic Device with an Integrated Freeze-Dried Cell-Free Protein Synthesis System for Small-Volume Biosensing

Microfluidic devices enable the precise operation of liquid samples in small volumes. This motivates why microfluidic devices have been applied to point-of-care (PoC) liquid biopsy. Among PoC liquid biopsy studies, some report diagnostic reagents being freeze-dried in such microfluidic devices. This type of PoC microfluidic device has distinct advantages, such as simplicity of the procedures, compared with other PoC devices using liquid-type diagnostic reagents. Despite the attractive characteristic, only diagnostic reagents based on the cloned enzyme donor immunoassay (CEDIA) have been freeze-dried in the microfluidic device. However, development of the PoC device based on the CEDIA method is time-consuming and labor-intensive. Here, we employed a molecule-responsive protein synthesis system as the diagnostic reagent to be freeze-dried in the microfluidic device. Such molecule-responsive protein synthesis has been well investigated in the field of molecular biology. Therefore, using the accumulated information, PoC devices can be efficiently developed. Thus, we developed a microfluidic device with an integrated freeze-dried molecule-responsive protein synthesis system. Using the developed device, we detected two types of bio-functional molecules (i.e., bacterial quorum sensing molecules and mercury ions) by injecting 1 µL of sample solution containing these molecules. We showed that the developed device is applicable for small-volume biosensing.