Team:Shanghai United HS/Safety

Shanghai_United_HS

Safety

To ensure the safety of the whole experiment, all of them have been done under supervision of instructors. A day before the experiment started, our whole group had a training session about the safety precautions in the laboratory. For instance, learning the correct usage of each lab equipment.

Laboratory safety precaution

1. Do not eat, drink, smoke, handle contact lenses, apply cosmetics, or store food for human consumption in the laboratory.
a. Only sealed water bottles are allowed in the laboratory area

2. Always wear appropriate personal protective equipment including but not limited to lab coats, gloves, etc
a. Change gloves when contaminated, and dispose of used gloves with other contaminated laboratory waste.

3. Wash your hands after working with potentially hazardous materials and before leaving the laboratory.

4. Label the container carefully and sophisticated to help the recognition of materials

Laboratory Environment

We’ve done all our experiments in a lab space provided by ATLATL. They’ve been trying to improve lab safety, through introducing many new products and technologies which have surfaced in the laboratory ventilation equipment industry in recent years. Our lab is classified as biosafety level 1(BSL-1), meaning that the lab minimizes the usage of hazardous chemical materials and the microbes used cause little to no risk to healthy individuals. Laboratory biosafety practices are based on the principle of containment of biological agents to prevent exposure to laboratory workers and the outside environment. This is done through wearing and leaving the lab coats only inside the lab, washing hands before exiting the lab, keeping all lab instruments sterilized and never leaving the laboratory with any contaminated equipment.

Biomaterial

The bacterial strain used throughout experiments are:
        1. Escherichia coli strain nissle 1917
        2. Escherichia coli DH5a
        3. Lactobacillus casei (To replace the pathogenic bacteria-Staphylococcus aureus in function tests)

Escherichia coli is a commonly used bacteria strain during biology experiments since it is quite safe to use compared to other bacterial kinds. The disposure of the bacteria is one of the major safety issues that need to be taken into consideration. Regarding the Lactobacillus casei, it is the probiotics in food industrial which performs little or nearly zero negative effect on human health.

Activator sequences needed for reconstructing plasmid:
ompA-ClyR-F: CTAACAGGAGGAATTAACCATGaaaaagacagctatcgcgattg
ompA-ClyR-R: CAAGCTGGAGACCGTTTAAACCACCTGCCGCGATGTttaatg

pBAD-F: cattaaACATCGCGGCAGGTGGTTTAAACGGTCTCCAGCTTG
pBAD-R: caatcgcgatagctgtctttttCATGGTTAATTCCTCCTGTTAG

BP_arab_F:tttCACCTGCGATCACGCATGAATGGGACAGTAAAAGTC (PaqC1_ACGC)
BP_arab_R:tttCACCTGCCGCGATGTTCAAGCCAGATCGACGGTCAAC (PaqC1_ACGC)

AmilGFP_F: tttCACCTGCGATCACGCatgtcttattcaaagcatggc (PaqC1_ACGC)
AmilGFP_R: tttCACCTGCCGCGATGTttatttaaccttcaaagggttaac (PaqC1_TACA)

The genetic cloning template:
Arabinofuranosidase and AmilGFP
Gene synthesis 1: pET-32a-ompA-PaqC1-ClyR-6His- PaqC1
Gene synthesis 2:pUC57-kan-mini-J23101-RBS-ompA- PaqC1- PaqC1-TT

Product

Since our product will be made into toothpaste, gum etc, safety issues are our priority. The vast majority of our experiment is based on bacteriophage lyase ClyR. Comparing to other products such as fluorine toothpaste, ClyR protein is more gentle, and has less side effect on symbiotic cells, which play a significant role in maintaining the eco balance amongst the oral bacterials.

Nucleic acid electrophoresis, testing for center genetic sequence, protein electrophoresis, antibacterial activity are used as characterization methods to ensure that the product is working perfectly.

Due to safety and ethical consideration, we will not perform any testing on animals nor humans. Without clinical trials, there is an undetermined risk of possible side effects of our product. There needs to be further research done regarding this specific issue to ensure the safety usage of the product.

References

1. “Chapter 4, Biosafety Manual: Biosafety Principles | Environmental Health & Safety.” University of Nevada, Reno, www.unr.edu/ehs/policies-manuals/biosafety-manual/chapter-4.

2. Sonnenborn, Ulrich, and Jürgen Schulze. “The Non-PathogenicEscherichia Colistrain Nissle 1917 – Features of a Versatile Probiotic.” Microbial Ecology in Health and Disease, vol. 21, no. 3-4, Jan. 2009, pp. 122–158, 10.3109/08910600903444267.

3. “Your Website Title.” Atlatl.cn, atlatl.cn/events/details?id=319. Accessed 14 Aug. 2021.