Time: 21:15-23:00
Location: Room 234, College of Life Sciences, Zhejiang University, China
Recorder:Li Jiachen
Highlights:meet-and-greet,the first meeting for ZJU-China IGEMers!
#1 Instructor's speech
#2 Team member self-introduction including wet lab group and dry lab group.
#3 Speech by the last captain: introduce and emphasize the attention.
#4 Specifying team rules
Futher plan:
Reading articles and journals.Everyone should do 1-2 self-designed projects and presnt in the next meeting.
Time: 21:15-23:45
Location:
Room 413, National Demonstration Center for Experiment Biology Dducation (ZJU), Zijin'gang Campus, Zhejiang University, China
Recorder: Gao Liaoliao
Highlights: Raise of the programs
#1 Detection of thyroid cancer by specific biological probe -By Pan Zicai
#2 Using biofilm to build a bioengineering platform of nano materials -By Han yibing
#3 Using biofilm to improve the enrichment and recovery efficiency of rare earth elements in e-waste -By Lin Hongying
#4 Using yeast and soybean protein to produce plant meat and improve ordinary vegetarian meat -By Li Jiachen
#5 Turing pattern -By Yan Zhijian
#6 Using algae to solve the treatment of tumor hypoxic area; Drug targeted delivery -By Su Yuyan
Future plan:
Narrow the scope of alternative projects, think about improvement of the programs and put forward new ideas.
Time: 21:15-23:30
Location:Room 413, National Demonstration Center for Experiment Biology Dducation (ZJU), Zijin'gang Campus, Zhejinag University, China
Recorder: Pan Zicai
Key summary: Brainstorming and sharing
#1: mold biocomputing --pass -By Yan Zhijian
#2 Make leather from fungi -By Ye Zihang
#3 Thyroid FNA testing -- what are the advantages of mab testing to be solved -By Pan Zicai
#4Artificial meat -By Li Jiachen
#5 Biofilm recovery of e-waste -By Lin Hongying
#6Drug delivery system for intestinal probiotics in the treatment of intestinal bleeding and targeting specific cells -By Su Yuyan:
#7 Ship protective coating made by biofilms, microbial fuel cell -By Han Yibing
#8 Cell factory synthesis of breast milk oligos accharides -By Liu Junyi
#9 Plant vaccines -By Ren Zhuoyun
#10 Bacterial imaging system -By Gao Liaoliao
Future plans
brainstorming
Time: 21:15-23:30
Location:Room413,National Demonstration Center for Experiment Biology Dducation(ZJU),Zijin'
Gang Campus, Zhejiang University, China
Recorder: Zihang Ye
Key summary: 3.13 Plan report and brainstorming sharing
#1 In vitro synthesis of FDCA, production of protein plastics -By SuYuyan,YeZihang
# 2Mercury-responsive promoter Pmer,amercaptogen-rich biofilms of Pseudo monassp -By LiJiachen,PanZicai
#3 Modification of nitrogen fixation with the gramineus symbiotic bacterium Klebsiellak.
variicola137 -By LiuJunyi,YanZhijian:
#4 Modified Bacillus subtilis for innovative application of pethair and keratin -By HanYibing,Lin
Hongying
#5 Accurate diagnosis of thyroid cancer -By PanZicai
#6 Antibiotic detection--CRISPR-12a, micro integral series -By LinHongying
#7 RPA technology combined with fluorescence quenching is relieved. Antibiotics interfere with RPA amplification and slow down the rate of fluorescence quenching -By Yan ZhiJian
Future plans
#1 Application and Innovation of keratin -By HanYibing
#2 Production of bio-plastics -By Su Yuyan, Ye Zihang
#3 Heavy Metal -By LiJiachen, PanZicai
Time: 21:15-23:30
Location: Room 413, National Demonstration Center for Experiment Biology Dducation (ZJU), Zijin’gang Campus, Zhejinag University, China
Recorder: Liu junyi
Highlights: Brain storm
#1 Secreting high-strength biological membrane -By Su Yuyan
#2 Multiple examination of soil pollution -By Li Jiachen
#3 Cell factory producing bio-plastics -By Ye Zihang
#4Modification of nitrogen-fixing bacterial for graminaceous crop; Engineered bactiria resolving hairball in pet’s stomach -By Liu Junyi
Future plan:
#1 bio-plastics -By Su Yuyan, Ye Zihang
#2 biological membrane -By Gao Liaoliao, Ren Zhuoyun
#3 soil pollution examination -By Li Jiachen, Pan Zicai
#4 nitrogen-fixing bacteria -By Liu Junyi, Yan Zhijian
#5 Resolving hairball -By Lin Hongying, Han Yibing
Time:21:15-23:00,March 21
Location:Room 413, National Demonstration Center for Experiment Biology Education (ZJU), Zijin'gang Campus,Zhejiang University, China
Recorder:Han Yibing
Highlights:
Summary: Wet Team members report on new ideas gathered this week
#1 Use gene knockout and agrobacterium infestation to solve the problem of crop incompatibility -By Yan Zhijian
#2 Temporarily hiding therapeutic bacteria outside the immune system allows them to treat rheumatoid arthritis more effectively -By Su Yuyan
#3 Producing self-healing cement from Bacillus -By Li JiaChen
#4 Hyphal material for the production of bio-plastics or insulating materials -By Li Jia-Chen
#5 Fibrosome of fusion catalysis, assembly, substrate binding, and cell binding modules -By Liu Jun-Yi
#6 Using gut microbes to treat depression -By Gao Liao-Liao
#7 Transform exophytic mycorhize -By Ye Zi Hang
#8 Magnetic protein -By Ren Zhuo Yun
Time: 21:15-23:15
Location:
Room 413, National Demonstration Center for Experiment Biology Education (ZJU), Zijin'gang Campus,Zhejiang University, China
Recorder: Zhuoyun Ren
Highlights: deep research on designed project
#1 The advantages and disadvantages of Bacillus subtilis as a chassis organism are presented -By Zicai Pan
#2 The application of synthetic organisms in restorative dentistry mentioned -By Yibing Han
#3 Complementary component designed -By Junyi Liu
Summary: Summarizes all the progress of the experiment from the initial design to today
Next steps: get feedback from different stakeholders and supplement our design according to the practical use
Time: 21:15-23:50
Location:
Room 413, National Demonstration Center for Experiment Biology Education (ZJU), Zijin'gang Campus,Zhejiang University, China
Recorder: Zhijian Yan
Highlights:
#1 Cement is more inclined to restoration of cultural relics, and its application in the field of architecture may not be very meaningful proposed -By Liaoliao GAO
#2 An innovative idea: Spirulina transformation system, using Spirulina to produce medical vaccines (can be made into oral vaccines) proposed By -Zihang Ye
#3 Utilize Bioconjugate to degrade corn stover, a new approach raised -By Zihang Ye
#4 Phage MS2 system detects the presence of specific sequence RNAs -By Zhijian Yan
#5 Brainstorm: 1) kraft paper to produce xylitol; 2) astaxanthin; 3) biological stapling; 4)pectin to jam; 5) plant terpane biosynthesis; 6) population sensing -By Zhuoyun Ren
#6 For Cd project: two possible pathways -By Zicai Pan, Hongying Lin
- Enrichment of cadmium by extracellular substrate-bound Pseudomonas aeruginosa
- Surface-based enrichment of cadmium by displaying MT protein
#7 For MICP project: research findings on specific synthetic biology design -By Junyi Liu, Yibing Han, Jiachen Li
- Related items/products are already available
- Use Bacillus alkalophilus as a chassis to sense pressure as a repair stop signal
- Expression of filamentous fibronectin to bind calcium carbonate, increase extracellular polysaccharide to improve crystal structure
- Culture material and calcium source can be used waste (calcium source: oyster shell, culture material: sesame seed residue (spore production), soybean meal (bacterium production))
Time: 21:15-23:00
Location:Room 413, National Demonstration Center for Experiment Biology Education (ZJU), Zijin'gang Campus, Zhejiang University, China
Recorder: Su Yuyan
Key summary: Brainstorming and sharing
#1: Develop own ACCBP components/ Promoter of stress perception -by Yan Zhijian
#2 Using microcapsule material packaging/Add alkali resistance system -by Li jiachen
#3 Riboswitch/find Nitrogen responsive promoter BBa_K216005 -by Ye zihang
#4 Summarize -by Su Yuyan
Using MICP principle to restore stone relics:
1. Chassis organism: Choose Bacillus subtilis
2. Metabolic pathways: ① Formation of calcium carbonate: the formation of calcium carbonate precipitation using amorphous calcium carbonate binding protein; Carbonic anhydrase was introduced to enhance the secretion of carbonate.
②The l-sucrose gum gene was introduced to synthesize the glue used for gluing calcium carbonate.
③ Secretedilamentous protein, as the supporting combined material.
3. Motor pathway: swrA and SFP genes of flagella were introduced to restore the motor ability of Bacillus subtilis and enable it to expand and grow on solids.
4. Other parts: alkali resistance, high salt resistance; Kill switch; Quorum sensing system.
5. Material preparation: soybean meal and other agricultural fertilizers are used as nutrient sources; Oyster shells were used as the source of calcium ions.
Time:21:30-23:00
Location:Room 413, National Demonstration Center for Experiment Biology
Dducation (ZJU), Zijin'gang Campus, Zhejinag University, China
Recorder:Lin Hongying
Highlights:The design of relic restoration project
#1 The introduction of the project from Newcastle in 2010, including germination part, swarming part, CaCO3 precipitation part, filamentous cell formation part and so on. ——By Li Jiachen
#2 The introduction of colloid parts and how to develop our own ACCBP parts and pressure-sensing parts. ——By Yan Zhijian
#3 The design of the safety part and quorum sensing part. ——By Han Yibing
#4 The introduction of the project to detect the concentration of N, P elements in polluted water. ——By Ye Zihang
Time: 21:15-23:00
Location: 413, National Demonstration Center for Experiment Biology Education (ZJU), Zijin'gang Campus, Zhejiang University, China.
Recorder:Li Jiachen
Highlights:we invite the team from the International Union College of Zhejiang University to talk about projects and change the suggestions of each other.
#1 Our project introduction
#2 International Union College of Zhejiang University team's project introduction
#3 question and answer
#4 discussion the problems and the solution
Future plan:
Further improve the design of the project, taking into account the water permeability of the stone, contamination by other bacteria, the resistance of bacteria, cost, hardware and so on.
Time: 21:15-23:00
Location:
Room 413, National Demonstration Center for Experiment Biology Dducation (ZJU), Zijin’gang Campus, Zhejinag University, China
Recorder: Liu junyi
Highlights: improvement of our current project
#1 Using silica gel as our padding -By Gao Liaoliao
#2 Using hypha to provide support frame -By Yan Zhijian
#3 Adding metal ion to prevent harmful bacterium -By Su Yuyan
#4 Bonding padding material by modified biological membrane -By Pan Zicai
Future plan:
Asking professors for the feasibility of the new designs; exploring new aspects for project improvement
Time:20:00-21:00
Location:Online Meeting through Dingtalk
Recorder:Lin Hongying
Highlights:Review of current project about relics restoration
Guest:Associate Professor Dr. Yulan Hu at Zhejiang University, whose main research interest is biotechnology and cultural relics protection.
#1 Evaluation:The project is theoretically feasible in the lab, but most stone relics are unmovable in the environment and other environmental factors like temperature and humidity should be considered.
#2 Detailed problems
1. The lack of oxygen in the deep
When the bacterium swimming into the deep cracks, their proliferation may be influenced due to the lack of oxygen affecting the repair effect.
2. The strength of the adhesive protein
The strength of the adhesive protein should be tested. And the protein strength will gradually reduce as time goes by.
3. The corrosion of cultural relics caused by microorganism.
The organic acids metabolized by microorganisms may cause corrosion to the stone relics, and we can introduce some microorganisms secreting alkaline substances.
4. The water content of the filler
Some stone relics can absorb the water in the medium so that our filler will become dry
5. How to modify our bacterium to gain a more competitive edge than other microorganisms in the cracks?
6. How do they evaluate the effect of the repair work?
We can evaluate the effect from some perspectives like safety, strength, et.al.
Time: 21:15-23:50
Location:
Room 413, National Demonstration Center for Experiment Biology Education (ZJU), Zijin'gang Campus,Zhejiang University, China
Recorder: Zhijian Yan
Highlights:
#1 Found parts in iGEM Website to design pathways from Sucrose to Fructans
#2 Analogue research in ACCBP
- How to design our own ACCBP module: Ca2+; Mg2+ aqueous solution
- Cloning of CARP protein from coral for precipitation of calcium carbonate under wide pH conditions
Summary: Some details in project design with different approaches
Time:21:15-23:00,April 27
Location:Room 413, National Demonstration Center for Experiment Biology Education (ZJU), Zijin'gang Campus,Zhejiang University, China
Recorder:Pan Zi-Cai
Highlights: Summarize the progress of HP and future planning, explain experiment arrangement and report on this week's progress.
#1 Hardware, construction for artifact crack detection (ultrasound & X-ray) -By Xie Yu-Shu
#2 Model, Ratio optimization of binders, strain growth relationships, screening of optimal quorum sensing systems using machine leaning-By Wang Yu-Kun
#3 Breeding strains gel to protect cultural relics-By Su Yu-Yan
#4 Using sulfate reducing bacteria to clean dirt on cultural relics-By Li Jia-Chen
#5 Improve immobilization efficiency by immobilizing carbonic anhydrase-By Lin Hong-Yin
#6 Produces bacterial cellulose, thus forming crosslink with other substances to form cleaners for rough surfaces-By Han Yi-Bin
#7 Rapid generation of materials with certain structural mechanics using Bacillus subtilis, organophosphate and barium alt systems-By Pan Zi-Cai
Future plan:
Carry out preliminary experiment.
Explore the surface cleaning scheme of cultural relics
Design quorum sensing systems
Construct ACCBP related modules
Explore new topics
Time: 21:15-23:33
Location:
Room 413, National Demonstration Center for Experiment Biology Education (ZJU), Zijin'gang Campus, Zhejiang University, China
Recorder: Gao Liaoliao
Highlights: Raise of the programs
#1 Bacterial targeted thrombolysis -By Gao Liaoliao
#2 Using RNAi to Kill nematodes -By Liu Junyi & Lin Hongying
#3 Measuring the expression level of endogenous mRNA by CRISPR system -By Yan Zhijian
#4 Treatment of triple negative breast cancer by simcell & Minisimcell -By Li Jiachen
#5 Fresh keeping of flowers - By Ren Zhuoyun
#6 Cleaning with stable bacteria strains - By Pan Zicai
#7 Biological cleaning, using paper gel bacteria to remove broad-spectrum stains - By Han Yibing
#8 Design of bacterial combined CaCO 3 scaffolds - By Ye Zihang
Future plan:
Cultural relics restoration, drug delivery system in cancer treatment, sewage detection capsule.
Time: 21:15-23:00
Location:Room 413, National Demonstration Center for Experiment Biology Education (ZJU), Zijin'gang Campus, Zhejiang University, China
Recorder: Han Yibing
Key summary: content introduction of iGEM opening ceremony and experimental schedule
# 1 Introduction to relevant contents of the opening ceremony (providing a slack to communicate with foreign iGEM teams and follow-up in-depth exchanges with China Agricultural University) -By Liu Junyi
Future plans:
#1 Experimental design of carbonic anhydrase -By Li Jiachen, Liu Junyi
#2 Experimental design of accbp mineralized peptide -By Ye Zihang, Lin Hongying, Su Yuyan
#3 Experimental design of high sugar suicide module -By Han Yibing, Ren zhuoyun, Yan Zhijian
#4 Experimental design of quorum sensing -By Gao Liaoliao, Pan Zicai
#5 Experimental design of measurement improvement -By Gao Liaoliao, Pan Zicai, Liu Junyi
#6 Write the experimental application -By Pan Zicai
#7 Docking ppt with Guo Donglai -By Han Yibing
Time:21:15-23:00
Location:Room413,National Demonstration Center for Experiment Biology Dducation(ZJU),Zijin'gang Campus,Zhejinag University,China
Recorder:YeZiHang
Key summary:rule explanation,experiment division
#1 Parts related matters interpretation -By Yan Zhijian
#2 Introduction to iGEM rules -By Su Yuyan
#3 A sensor literature explanation -By Yan Zhijian
#4 Grouting method; Modeling based on fracture conditions and media fluidity -By Lin Hongying
Future plans:
#1 Lin Hongying:Responsible for safety certification
#2 Su Yuyan:Contact Teacher Wang Baojun
#3 Su Yuyan,Yan Zhijian:Experimental design of cultural relic removal
#4 Jiachen Li,Junyi Liu: Carbonian hydrase partial experimental design
#5 ZihangYe,HongyingLin:ACCBP part experimental design
#6 HanYibing,YanZhijian: Experimental design of part of suicide module
#7 Gao Liaoliao,PanZicai: Experimental design of quorum sensing
#8 WangYu kun: Modeling based on fracture conditions and media fluidity
- Public
Purify a strain of WB600 by plate streaking from commercialize B.subtilis strain.
Prepare culture media containing Tetracycline and test Shuttle plasmid pHY300PLK
Get the GFP and mRFP sequence from distribution plate.
- Public
Separately construct pHY300PLK plasmids with GFP and RFP, in order to test transforming conditions of B.subtilis.
- Public
Separately construct pHY300PLK plasmids with GFP and RFP, in order to test transforming conditions of B.subtilis.
- Part Measurement
Amplifate luciferae, luciferin-regenerating enzyme, and separately construct three pet28(a)+ plasmids:
(a) pet28a(+) with luciferase,(b) pet28a(+) with luciferase regenerating enzyme,(c) pet28a(+) with both luciferase and luciferin-regenerating enzyme.
Transform three plasmids to E.coli BL21
- Public
Try electric transform of B.subtilis but failed
- Biological scafforld
Synthesis plasmids PHY300PLK-HagT209C-SpyT , PHY300PLK-EutM-SpyC , PET-28(a)- spyT-mRFP , PET-28(a)-spyC-mRFP by Genscript .
- Quorum Sensing Part
construction of plasmids: Oxylimited-GFP-terminator
- Biological scafforld
Transform PET-28(a)- spyT-mRFP , PET-28(a)- spyT-mRFP in BL21
Explore the protein inducing condition
Complete the induction of spyT-mRFP and spyT-mRFP
- Biosafety
Improve the 2012 LMU's Bacillus subtilis genome extraction protocol: only chloroform isoamyl alcohol (24:1) is used to extract protein, and the extraction time of Bacillus subtilis genome DNA is shortened to within 2h.
spaBCTS and PsacB sequences have been successfully cloned from genomic DNA.
Improvement of protein SDS-PAGE process: a new protein glue dyeing solution was developed.
- Calcium Carbonate Production part
Do codon optimization for Carbonic Anhydrase(CA) and construct the plasmid PHY300PLK-P43-CA1
Plasmids PHY300PLK-P43-CA1 and PHY300PLK-ACCBP synthesis (by company)
- Quorum Sensing Part
Construction of plasmids: Oxylimited-GFP-terminator
- Biological scafforld
Explore the proprior condition for the HA-tag protein IP purification
- Biosafety
PHY-PSacB-GFP-Terminator plasmid and PHY-Pveg-PsacB-GFP-Terminator plasmid were constructed successfully.
- Calcium Carbonate Production part
Transform PHY300PLK-P43-CA1 in BL21
PHY300PLK-P43-CA1 plasmid extraction
- Public
Several attempts to electrotransform Bacillus subtilis failed.
We have done a successful attempt at chemical transformation of B.subtilis
- Quorum Sensing Part
construction of plasmids signal-GFP, and Characterization in BL2; Characterization of immunity part in BL21;explore the effect of Bacillus subtilis inhibitory concentration
- Biological scafforld
Explore the chemical confromation method of Bacillus subtilis .
Explore the protein expression condition of Bacillus subtilis .
- Biosafety
Try to refer to the protocol of LMU to chemically transform Bacillus subtilis, and succeed.
PHY-PsacB-Pveg-GFP-Terminator plasmid was characterized in Bacillus subtilis. It had no induction activity, but had expression activity
PHY-PsacB-GFP-Terminator plasmid was characterized in Bacillus subtilis, with no induction activity and suspected weak expression activity
- Calcium Carbonate Production part
PHY300PLK-P43-CA1 electrotransformation to WB600 (failed)
We have done a successful attempt at chemical transformation of B.subtilis
- Quorum Sensing Part
Confirm the function of immunity part in BL21.
- Biological scafforld
Purify the spyC-mRFP (with 6*His tag) by His-tag Purification Resin
Purify the spyT-mRFP (with HA tag) by Anti-HA Magnetic Beads
- Biosafety
The concentration of sucrose (1% - 10% sucrose) was changed to characterize the induction effect of PsacB plasmid. It was found that it had only weak expression activity and no induction activity. It was considered to optimize the promoter to reduce leakage expression.
Transforming PHY psacb MazE / F into E. coli and extracting plasmid
- Calcium Carbonate Production part
Restructuring plasmid (add Terminater7 to PHY300PLK-P43-CA1)
- Quorum Sensing Part
Transform plasmids into WB600, including Oxylimited-GFP-terminator, Oxylimited-SpaBCTS-terminator,signal-GFP. Run SDS-Page to confirm the phenotype. However, failed.
- Biological scafforld
Construct the PHY-PsacB-EutM-spyC-terminator (add the promotor and terminator)
- Biosafety
The plasmid was reconstructed and the reporter gene was inserted into the HindIII digestion site (shortening the distance between the start codon and RBS) to try to improve the expression efficiency
- Calcium Carbonate Production part
Construct the plasmid PHY300PLK-P43-CA2-T1-T7(from Parts: BBa_K2232014)
- Quorum Sensing Part
Construction of plasmids: P43-GFP,P43-SpaBCTS, and confirm the phenotype in BL21 using 0.1mg/ml Bacillus subtilis to trigger the function of signal-GFP plasmid in WB600
- Biological scafforld
Induce the expression of EutM-SpyC in Bacillus subtilis .
Explore the experimental protocol of EutM-SpyC
Construct the PET-28(a)-EutM-spyC plasmid
- Biosafety
The characterization could not be completed due to the dispersion of SDS-PAGE small molecule proteins.
Try to improve the efficiency of promoter induced expression
replacement of reporter gene BBa_K1159001(FirelyLuciferase) BBa_I712019(NanoLuciferase) add RBS (Spovg-RBS, available before Pveg)
- Calcium Carbonate Production part
PHY300PLK-P43-CA2-T1-T7 chemical transformation to WB600. Cultivating the engineered WB600 in LB (Tc+) for 24hExpression of carbonic anhydrase in Bacillus subtilis
Construct the PET-28(a)-ACCBP plasmid
- Quorum Sensing Part
Purify the plasmid and transform plasmids into WB600. Confirm the phenotype of P43-GFP plasmid in WB600 by SDS-Page.
- Biological scafforld
Construct the PHY-P43-HagT209C-SpyT (add the promotor and terminator)
Complete the western blotting verification of spy system
- Biosafety
Optimize promoter recombinant full-length Pveg-sacB (linearized vector with homologous arm and Pveg have been prepared).
Recombinant PVG leader RNA (recombine the constitutive promoter PVG and the leader RNA upstream of sacB gene).
- Calcium Carbonate Production part
Transforme the PET-28a(+)-ACCBP into the BL21.
- Quorum Sensing Part
Confirm the GFP expression triggered by 0.1 mg/ml Bacillus subtilis inWB600 and run SDS-Page.
- Biological scafforld
Transform PET-28(a)-EutM-SpyC in BL21
Induce the EutM-SpyC and purify the EutM-SpyC(with 6*His tag ) by His-tag Purification Resin
Verify the function of protein
- Biosafety
Construct plasmids, but they have been failing.
- Calcium Carbonate Production part
Purification of carbonic anhydrase and SDS-PAGE electrophoresis
Detection of esterase activity of carbonic anhydrase
- Quorum Sensing Part
Construction of plasmids: Oxylimited-SpaBCTS-terminator-New. Transform plasmids into WB600.
- Biological scafforld
Construct the PHY-P43-HagT209C::SpyTag588-DT plasmid by overlapping PCR
- Biosafety
- The PHY-PsacB-FierlyLuciferase-Terminator plasmid, PHY-PsacB-NanoLuciferase-Terminatorplasmid, and PHY-Pveg-LR-spoVGRBS plasmid were successfully constructed.
- Calcium Carbonate Production part
Cultivating the engineered WB600 in SMM (Tc+) for 48h
- Biological scafforld
Induce the expression of EutM-SpyC in E.coli .
- Calcium Carbonate Production part
Determination of the ability of Carbonic Anhydrase to catalyze the hydration of CO2
Veify the ACCBP expression in the E.coli
- Biological scafforld
Purify the EutM-SpyC from E.coli and utilize the TEM to detect the morphology of EutM.
Transform the PHY-Hag into the Bacillus subtilis.
- Calcium Carbonate Production part
Determination the ability of Carbonic Anhydrase to catalyze the precipitation of CaCO3
Explore the expression condition of ACCBP in the E.coli
- Calcium Carbonate Production part
SEM exploring the crystal structure of precipitated amorphous calcium carbonate
Explore the expression condition of ACCBP in the E.coli
Construct the PET-28a(+)-Δ16 ACCBP plasmid
- Biological scafforld
Transform the plasmid PHY-P43-Hag into the Bacillus subtilis
Verify the function of Hag
Verify the feasibility of biological scaffold
- Biosafety
Transform pHY300PLK containing BBa_K4202045(PsacB-mazE-mazF), BBa_K4202033(Pveg-1xLR-GFP), BBa_K4202034(Pveg-2xLR-GFP), BBa_K4202035(Pveg-3xLR-GFP), and BBa_K4202036(Pveg-1xLR-GFP) into Bacillus subtilis WB600 strain.
Characterized the above parts in media containing different sucrose concentrations. The growth curve was measured using Nanodrop (only in the characterization of BBa_K4202045) and the fluorescence intensity was measured using flow cytometer.
- Calcium Carbonate Production part
ACCBP was expressed and purified according to the conditions obtained in the pre-experiment
Protein structure and function verification of ACCBP.
