"
The beast and its Achilles heel:
A novel target to fight multi-resistant pathogenic bacteria
Download as pdf
Basic Safety Questions
1. Please describe the chassis organism(s) you will be using for this project. lf you will be using more than one chassis organism, provide information on each of them:
|
Species |
Strain no/name |
Risk Group |
Risk group source link |
Disease risk to
humans? If so, which disease? |
|
E.coli (K12) |
XL1 blue |
1 |
www.absa.org/riskgroups/bacteria search.php?genus=&species=coli |
Yes. May cause
irritation to skin, eyes, and respiratory tract, may affect kidneys. |
|
E.coli (K12) |
DH5 alpha |
1 |
www.absa.org/riskgroups/bacteria search.php?genus=&species=coli |
Yes. May cause
irritation to skin, eyes, and respiratory tract, may affect kidneys. |
|
E.coli(B) |
T7 express(NEB) |
1 |
www.absa.org/riskgroups/bacteria search.php?genus=&species=coli |
Yes. May cause
irritation to skin, eyes, and respiratory tract, may affect kidneys. |
|
E.coli(B) |
BL21 |
1 |
www.absa.org/riskgroups/bacteria search.php?genus=&species=coli |
Yes. May cause
irritation to skin, eyes, and respiratory tract, may affect kidneys. |
2. Highest Risk Group Listed:
Risk group 1
3. List and describe all new or modified coding regions you will be using in your project. (If you use parts from the 2013 iGEM Distribution without modifying them, you do not need to list those parts.)
|
Part number. |
Where did you get the physical DNA for this
part (which lab,synthesis company, etc) |
What species does this part originally come
from? |
What is the Risk Group of the species? |
What is the function of this part, in its
parent species? |
|
BBa_K1045000 |
Cloned from purchased Mycobacteria
genome |
Mycobacterium segmatis |
2 |
The operator DNA sequence that DarR binds |
|
BBa_K1045001 |
Cloned from purchased Mycobacteria
genome |
Mycobacterium segmatis |
2 |
Code regulatory protein DarR,
which binds c-di-AMP |
|
BBa_K1045002 |
Cloned from Bacillus genome |
Bacillus subtilis |
1 |
The c-di-AMP
sensing riboswitch YdaO |
|
BBa_K1045003 |
Cloned from Listeria monocytogenes genome |
Listeria monocytogenes |
2 |
Diadenylate cyclase domain of L. monocytogenes,
cdaA |
|
BBa_K1045004 |
Cloned from the Bacillus substilis genome |
Bacillus substilis |
1 |
YdaO
Riboswitch with native Promoter and RBS |
|
BBa_K1045005 |
Cloned from the Bacillus substilis genome |
Bacillus substilis |
1 |
YdaO
Riboswitch with native RBS |
|
BBa_K1045006 |
Cloned from the Bacillus substilis genome |
Bacillus substilis |
1 |
YdaO
Riboswitch with native Promoter |
|
BBa_K1045007 |
Cloned from the Bacillus substilis genome |
Bacillus substilis |
1 |
YdaO
Riboswitch only |
|
BBa_K1045009 |
From 2013 distribution kit |
E.coli |
1 |
Terminator |
|
BBa_K1045010 |
From 2013 distribution kit |
E.coli |
1 |
Ribsome
binding site |
|
BBa_K1045011 |
From 2013 distribution kit |
E.coli |
1 |
Promoter |
|
BBa_K1045012 |
Assembled with BBa_K1045000 and BBa_J23110 |
|||
|
BBa_K1045013 |
Aseembled
with BBa_K1045012 and BBa_E0240 |
|||
|
BBa_K1045014 |
Assembled with BBa_K1045011 and
BBa_K1045013, |
|||
|
BBa_K1045015 |
Assembled with BBa_K1045014, BBa_K1045010 and
BBa_E0240 |
|||
|
BBa_K1045016 |
Assembled with BBa_K1045001 and
BBa_K1045009, |
|||
|
BBa_K1045017 |
Assembled with BBa_K1045015 and
BBa_K1045016 |
4. Do the biological materials used in your lab work pose any of the following risks? Please describe.
a. Risks to the safety and health of team members or others working in the lab?
The organism we used in the lab E.coli (K12) DH5 alpha, could be pathogenicto human beings. lt may cause irritation to skin, eyes, and respiratorytract and affect kidneys.
b. Risks to the safety and health of the general public, if released by design or by accident?
The E.coli strain we used do have pathogenicity to human. But the relatively low pathogenicity, short life and vulnerability to environment factors ofthe used strain have limited the risk to general public.
c. Risks to the environment, if released by design or by accident?
The E.coli strains we used during the project are transformed with plasmids containing antibiotic markers The release of those transformed strains may cause problems like the leakage of resistance genes to wild bacterial flora.
d. Risks to security through malicious misuse by individuals, groups, or countries?
The strains we used during our project do not contain hazardous genetic compartments. The risk through malicious misuse is quite low
5. If your project moved from a small-scale lab study to become widely used as a commercial/industrial product, what new risks might arise? (Consider the different categories of risks that are listed in parts a-d of the previous question.) Also, what risks might arise if the knowledge you generate or the methods you develop became widely available? (Note: This is meant to be a somewhat open-ended discussion question.)
The system we are trying to build is mainly meant for drug screening in pharmaceutical industry. The scale of production will always be limited.
6. Does your project include any design features to address safety risks? (For example: kill switches, auxotrophic chassis, etc.) Note that including such features is not mandatory to participate in iGEM, but many groups choose to include them.
No. But as the most used model organism in molecular biology, the risk of E.coli is relatively low.
7. What safety training have you received (or plan to receive in the future)? Provide a brief description, and a link to your institution’s safety training requirements, if available.
Before the project started, we all received instructions concerning the safety problems, such as how to protect ourselves in the lab, how to handle the biological materials properly to lower the risk to public and environment.
8. Under what biosafety provisions will / do you work?
a. Please provide a link to your institution biosafety guidelines.
www.uni-goettingen.de/en/401.html
(this website has links to all safety concerning documents, but all in german)
b. Does your institution have an Institutional Biosafety Committee, or an equivalent group? If yes, have you discussed your project with them? Describe any concerns they raised with your project, and any changes you made to your project planbased on their review.
www.uni-goettingen.de/de/401.html
c. Does your country have national biosafety regulations or guidelines? If so, please provide a link to these regulations or guidelines if possible.
d. According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab? (Check the summary table on page 3, and the fuller description that starts on page 9.) If your lab does not fit neatly into category 1, 2, 3, or 4, please describe its safety features [see 2013.igem.org/Safety for help].
The biosafety level of our lab is Categroy 1.
e. What is the Risk Group of your chassis organism(s), as you stated in question 1? If it does not match the BSL rating of your laboratory, please explain what additional safety measures you are taking.
Risk Group 1.
Faculty Advisor Name:
Jörg Stülke
Satefy Questions Part 2
