Team:HSiTAIWAN/Project/Framework

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　Oh hello there,I am Charlie,a trusty member of the "Herb Tasters" and also the brainiest E. coli in the colony.
　I know all the secrets of Chinese herbs and their magical healing powers.
　If you are up to a challenge,find me at team HSiTW at the jamboree.
　I am the one in a straw hat,showing them pearls.I will be waiting.
　Muhahahahaha

　Hi there! My name is Nu Zhen Chi. This is how I look like.
　　Take a closer look; guess which part of me is used as medicine?
(1) the root
(2) the stem
(3) the leaf
(4) the seed

　Ans.(4) the seed
　Name: 女貞子 (Nu Zhen Chi)
　Botanical Name: Ligustrum lucidum Aiton
　I can treat people who are yin deficient, and liver problems that cause dizziness,cataract of the eyes,
lower back pain, premature graying of the hair and tinnitus.

　Hello! My name is Chuan Xiong. This is how I look like.
　Make a guess, which part of me is used as medicine?
(1) the root
(2) the stem
(3) the leaf
(4) the seed

　Ans.(1) the root
　Name: 川芎 (Chuan Xiong)
　Botanical Name: Ligusticum chuanxiong Hort
　I help with blood regulation to prevent relevant to blood stasis and non-stop bleeding.I can also strengthen your qi circulation.
　In addition, I relieve you of physical pain, such as headaches, abdominal aches, chest pain, and muscle pain.
　Finally, I free the ladies of menstrual disorders and amenorrhea.

　What’s up? My name is Dang Gui. I can:
(1) stop coughing
(2) regulate mense
(3) reduce internal heat

　Ans.(2) Regulate mense
　Name: 當歸 (Dang Gui)
　Botanical Name: Angelica sinensis (Oliv.) Diels
　I can remove blood stasis and clots, so I am usually used to regulate menses,lubricate intestines to correct constipation, reduce swelling, expel pus.

　　Reference
　臧堃堂 (2005) 中華材輕百科-現代版本草綱目，山岳文化出版社，台北
　Non-Profit Organization Brion Research Institute of Taiwan.
　Chinese Herb Gallery. Jade Institute
　Herbal Glossary. Shen-Nong- Chinese Traditional Medicine
　　Acknowledgement
　Thank you for Non-Profit Organization Brion Resaerch Institute of Taiwan that provide us Chinese herbs and photos.

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  • Framework

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    Framework

    　　

    The goal of our Herb Tasters project is to design biosensors to detect heavy metals and aflatoxin in Chinese medicine. We used synthetic biology technology to make E. coli carry different types of plasmid which can express fluorescence in the presence of metal ions or aflatoxin. The prototype of our product is designed in a form of bio-kit package consisting of these biosensors. Also, we designed our product have biosafety mechanism to prevent of E. coli from spreading and polluting the environment. We conducted experiments to test the functions of biosensors and the effectiveness of biosafety mechanism

    Figure. Framework of The Herb Tasters

    Biosensor Function

    We designed new parts or used the previously existing parts to detect aflatoxin and three kinds of heavy metals—lead, copper, arsenic ions in Chinese medicine.

    1. Aflatoxin: we used aflatoxin metabolism and SOS response to design the circuit. Based on our literature review, we submitted five new basic parts for aflatoxin metabolism and we redo ligation for two composite parts

    2. Lead ion: the circuit consists of two parts: generator of the regulator PbrR which regulates the lead resistance operon; lead ion reporter of which the GFP express fluorescence when the dimer of lead ion and PbrR binds onto the PbrR promoter.

    For the lead biosensor, we designed an autoregulation mechanism to control the PbrR generation because the GFP expression will be affected when PbrR concentration is higher than lead lion. We submitted three new composite parts.

    3. Copper ion: we tried a new application of the device of the copa promoter with a GFP reporter (BBa_K1555000 ; iGEM14_NJAU_China), to detect Cooper ion in Chinese medicine.

    4. Arsenic ion: we tried a new application of GFP generator under Ars promoter (BBa_K1106004 ; iGEM13_Buenos_Aires) to detect the arsenic ions in the Chinese medicine.

    Functional Assays

    We conducted two sets of experiments: we tested the survival of the E.coli in Chinese medicine to test the false negative problem (make sure the no expression of GFP is due to E. coli death). We tested the biosensor function through measuring fluorescence in different ion concentration.

    Function Assays in Chinese Medicine

    We redo all experiments in different Chinese medicine. In addition, considering Chinese herbal medicines are plants and some heavy metal ions are essential micronutrients for their metabolism, we also use EDTA to chelate heavy metal ions containing in our testing Chinese herbs to accurately assess levels of heavy metal among these Chinese medicines.

    Modeling

    For the lead biosensor, we designed an autoregulation mechanism to control the PbrR generation because the GFP expression will be affected when PbrR concentration is higher than lead lion. We compared the GFP expression of auto-regulated PbrR generator and the generator under control with a constitutive promoter to model the effect of autoregulation.

    Product Biosafety Test: sterilization

    We have three steps of biosafety mechanism for our prototype. In one step, we used a special plant (hibiscus) to kill E. coli. Our experiment shows that hibiscus has a great function of bacteria sterilization.