Regulatory

Part:BBa_K2796036

Designed by: Shuangshuang Pu   Group: iGEM18_LZU-CHINA   (2018-10-04)


pTRE

Tet-induced regulatory expression system is a regulatory system for inducing gene expression based on the Tet-resistant operon specific to the transposon of Tn10 in E. coli.

Modeling

To activate our tetracycline-induced promoter, we administered tetracycline intravenously. However, excessive tetracycline can affect gastrointestinal tract, liver, blood system, central nervous system and kidney to different degrees. Tetracycline can also bind with phosphate in dentin and enamel, leading to teeth becoming yellow and bone dysplasia. Therefore, we need to establish a drug metabolism model to analyze and calculate the optimal drug concentration range during intravenous injection, to reduce the impact of tetracycline administration and provide certain mathematical basis for the selection of drug injection time and drug injection amount in practical application.

2018 LZU-CHINA 36 mode1.png

a.The metabolism of tetracycline in the stomach conforms to the rapid perfusion model.

2018 LZU-CHINA 36 mode2.png
2018 LZU-CHINA 36 mode3.png

Over time, tetracycline concentrations gradually drop in the blood and enter the stomach tissue. It was predicted that tetracycline in the stomach reached its peak at 10hours, and then its concentration gradually decreased with the degradation of tetracycline.

2018 LZU-CHINA 36 mode4.png

This diagram shows the functional relationship between tetracycline and absorption time and absorption coefficient in gastric tissue. The tetracycline concentration decreases gradually with the time of absorption, and the absorption efficiency is inversely proportional to the tetracycline concentration in the gastric tissue.
b.when we give the drug intravenously, the change of blood drug concentration conforms to the following formula:

2018 LZU-CHINA 36 mode5.png

K_a is based on a Nomograph designed by Franke et al. f=Q_M72/Q_b (Q_b is the dosage of the solution), the value of K_0=f K，V_d=clr/K_a ，K_a，f，K_0，V_d refer to the previous literature.

2018 LZU-CHINA 36 mode7.png

With the increase of time, tetracycline concentration in blood medicine gradually increased until reaching the plateau stage.

Experimental result

We successfully amplified pTRE from pTRIPZ plasmid and built it into pSB1C3.

2018 LZU-CHINA 36 gel.png

Reference

( Barr, W. H., Gerbracht, L. M., Letcher, K., Plaut, M., & Strahl, N. (1972). Assessment of the biologic availability of tetracycline products in man. Clinical Pharmacology & Therapeutics, 13(1), 97–108.)


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 271
  • 1000
    COMPATIBLE WITH RFC[1000]


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