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
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.
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)
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
ompA-ClyR-F: CTAACAGGAGGAATTAACCATGaaaaagacagctatcgcgattg
ompA-ClyR-R: CAAGCTGGAGACCGTTTAAACCACCTGCCGCGATGTttaatg
pBAD-F: cattaaACATCGCGGCAGGTGGTTTAAACGGTCTCCAGCTTG
pBAD-R: caatcgcgatagctgtctttttCATGGTTAATTCCTCCTGTTAG
pBAD-R: caatcgcgatagctgtctttttCATGGTTAATTCCTCCTGTTAG
BP_arab_F:tttCACCTGCGATCACGCATGAATGGGACAGTAAAAGTC (PaqC1_ACGC)
BP_arab_R:tttCACCTGCCGCGATGTTCAAGCCAGATCGACGGTCAAC (PaqC1_ACGC)
BP_arab_R:tttCACCTGCCGCGATGTTCAAGCCAGATCGACGGTCAAC (PaqC1_ACGC)
AmilGFP_F: tttCACCTGCGATCACGCatgtcttattcaaagcatggc (PaqC1_ACGC)
AmilGFP_R: tttCACCTGCCGCGATGTttatttaaccttcaaagggttaac (PaqC1_TACA)
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
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.