Limits...
Systematic approach to Escherichia coli cell population control using a genetic lysis circuit.

Hsu CY, Yu TC, Lin LJ, Hu RH, Chen BS - BMC Syst Biol (2014)

Bottom Line: Finally, according to design specifications, a systematic design approach is proposed to provide synthetic biologists with a prescribed I/O response by selecting proper promoter-RBS component set in combination with suitable inducer concentrations, within a feasible range.This study provides an important systematic design method for the development of next-generation synthetic gene circuits, from component library construction to genetic circuit assembly.In future, when libraries are more complete, more precise cell density control can be achieved.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Background: Cell population control allows for the maintenance of a specific cell population density. In this study, we use lysis gene BBa_K117000 from the Registry of Standard Biological Parts, formed by MIT, to lyse Escherichia coli (E. coli). The lysis gene is regulated by a synthetic genetic lysis circuit, using an inducer-regulated promoter-RBS component. To make the design more easily, it is necessary to provide a systematic approach for a genetic lysis circuit to achieve control of cell population density.

Results: Firstly, the lytic ability of the constructed genetic lysis circuit is described by the relationship between the promoter-RBS components and inducer concentration in a steady state model. Then, three types of promoter-RBS libraries are established. Finally, according to design specifications, a systematic design approach is proposed to provide synthetic biologists with a prescribed I/O response by selecting proper promoter-RBS component set in combination with suitable inducer concentrations, within a feasible range.

Conclusion: This study provides an important systematic design method for the development of next-generation synthetic gene circuits, from component library construction to genetic circuit assembly. In future, when libraries are more complete, more precise cell density control can be achieved.

Show MeSH

Related in: MedlinePlus

Inducible LuxR-regulated circuit with lysis gene in E. coli. A constitutive promoter continuously produces the activator protein LuxR. The protein LuxR needs to form a complex with the inducer AHL. The LuxR-AHL complex constitutes a quorum sensing mechanism. It activates the downstream promoter and enhances expression of the lysis gene when this complex accumulates.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4305986&req=5

Figure 2: Inducible LuxR-regulated circuit with lysis gene in E. coli. A constitutive promoter continuously produces the activator protein LuxR. The protein LuxR needs to form a complex with the inducer AHL. The LuxR-AHL complex constitutes a quorum sensing mechanism. It activates the downstream promoter and enhances expression of the lysis gene when this complex accumulates.

Mentions: For the convenience of description and explanation, as shown in Figure 2, a genetic lysis circuit is assembled by selecting a set of promoter-RBS components, namely, a constitutive promoter-RBS component Ci from Additional file 1 and an activator-regulated promoter-RBS component Am from Additional file 1. The lysis gene is embedded downstream of the activator-regulated promoter-RBS component. The genetic lysis circuit is divided into two stages. The first stage involves a constitutive promoter-RBS component Ci for producing regulatory protein, LuxR. The second stage involves an activator-regulated promoter-RBS component Am for driving the expression of the lysis protein. The external inducer AHL is used to regulate the lysis activity. The desired population response Nref(I) to different inducer concentrations described as follows:


Systematic approach to Escherichia coli cell population control using a genetic lysis circuit.

Hsu CY, Yu TC, Lin LJ, Hu RH, Chen BS - BMC Syst Biol (2014)

Inducible LuxR-regulated circuit with lysis gene in E. coli. A constitutive promoter continuously produces the activator protein LuxR. The protein LuxR needs to form a complex with the inducer AHL. The LuxR-AHL complex constitutes a quorum sensing mechanism. It activates the downstream promoter and enhances expression of the lysis gene when this complex accumulates.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4305986&req=5

Figure 2: Inducible LuxR-regulated circuit with lysis gene in E. coli. A constitutive promoter continuously produces the activator protein LuxR. The protein LuxR needs to form a complex with the inducer AHL. The LuxR-AHL complex constitutes a quorum sensing mechanism. It activates the downstream promoter and enhances expression of the lysis gene when this complex accumulates.
Mentions: For the convenience of description and explanation, as shown in Figure 2, a genetic lysis circuit is assembled by selecting a set of promoter-RBS components, namely, a constitutive promoter-RBS component Ci from Additional file 1 and an activator-regulated promoter-RBS component Am from Additional file 1. The lysis gene is embedded downstream of the activator-regulated promoter-RBS component. The genetic lysis circuit is divided into two stages. The first stage involves a constitutive promoter-RBS component Ci for producing regulatory protein, LuxR. The second stage involves an activator-regulated promoter-RBS component Am for driving the expression of the lysis protein. The external inducer AHL is used to regulate the lysis activity. The desired population response Nref(I) to different inducer concentrations described as follows:

Bottom Line: Finally, according to design specifications, a systematic design approach is proposed to provide synthetic biologists with a prescribed I/O response by selecting proper promoter-RBS component set in combination with suitable inducer concentrations, within a feasible range.This study provides an important systematic design method for the development of next-generation synthetic gene circuits, from component library construction to genetic circuit assembly.In future, when libraries are more complete, more precise cell density control can be achieved.

View Article: PubMed Central - HTML - PubMed

ABSTRACT

Background: Cell population control allows for the maintenance of a specific cell population density. In this study, we use lysis gene BBa_K117000 from the Registry of Standard Biological Parts, formed by MIT, to lyse Escherichia coli (E. coli). The lysis gene is regulated by a synthetic genetic lysis circuit, using an inducer-regulated promoter-RBS component. To make the design more easily, it is necessary to provide a systematic approach for a genetic lysis circuit to achieve control of cell population density.

Results: Firstly, the lytic ability of the constructed genetic lysis circuit is described by the relationship between the promoter-RBS components and inducer concentration in a steady state model. Then, three types of promoter-RBS libraries are established. Finally, according to design specifications, a systematic design approach is proposed to provide synthetic biologists with a prescribed I/O response by selecting proper promoter-RBS component set in combination with suitable inducer concentrations, within a feasible range.

Conclusion: This study provides an important systematic design method for the development of next-generation synthetic gene circuits, from component library construction to genetic circuit assembly. In future, when libraries are more complete, more precise cell density control can be achieved.

Show MeSH
Related in: MedlinePlus