Part:BBa_K1031441

J23106-HcaR

Characterization

In order to fine-tune the performance of HcaR biosensor, we constructed a library of Pc constitutive promoters at different intensity. BBa_K1031441 is composed of three elements, the constitutive Pc promoter J23106[1], coding sequence of HcaR and terminator B0015[2]. (Fig 1)

Fig 1 Construction of biosensor circuit Ph/J23106-HcaR

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 892
Illegal AgeI site found at 565
1000
COMPATIBLE WITH RFC[1000]

Data shown

Induction ratio of HcaR biosensor library test that utilizes different Pc promoters to control the expression of HcaR. The dashed box refers to data for Ph/J23106-HcaR biosensor circuit (Fig 2). This figure shows the biosensor circuit adopting J23106 obtains optimal performance.

Fig 2 Induction ratio of HcaR biosensor library test. Horizontal axis stands for different HcaR biosensor with Pc promoters of different strength. The expression strength of these constitutive promoters, J23113, J23109, J23114, J23105, J23106 is 21, 106, 256, 623, and 1185, respectively, according to the Partsregistry. Four kinds of aromatics shown with different color intensities, were tested following Test Protocol 1 [1]. Vertical axis represents the ON-OFF induction ratio. The dashed box refers to performance for Ph/J23106-HcaR biosensor circuit.

Various inducers were used to test dose-response curve of Ph/J23106-HcaR at the concentration at 10µM, 30µM, 100µM, 300µM and 1000µM (Fig 3)

Fig 3 Dose response curve of biosensor circuit Ph/J23106-HcaR. X-axis stands for concentration gradient of inducers at 10µM, 30µM, 100µM, 300µM and 1000µM. Different colors represent different kinds of inducers. Y-axis shows induction ratio reflected via fluorescence intensity.

biology
efficiency