Week1    27/10/2017

First meeting, Team FJNU-China was established.

Week2    03/2-10/2

Learn the molecular biology experimental techniques under the guidance of Dr.Yang.

Week3-Week7    Apr

Sent d-ldh, Tyrb and rocG to the company for synthesis.
Obtained d-ldh, Tyrb and rocG fragment by PCR.[Result1] [Result2][Result3]

Week8-Week12    May

1.Extract genome of E. coli Bw25113.
2.Construct d-ldh, Tyrb and rocG by Gibson assembly.
3.PCR verification.
[Result]

Week13-Week17    June

1.Extracted plasmid of E. coli BM4R
2.Cultured Clostridium acetobutylicum
3.Induction culture of engineering bacteria
4.SDS-PAGE verifies protein expression
[Result]

Week18    1/7-7/7

1.Synthetic primer for our experiments.
2.Cultured staphylococcus epidermidis.
3.Did a PCR pre-experiment to amplify atoD, atoA
4.Verified that if E.coli can grow without salt.
5.Held a summer camp with Quanzhou 7th High school and Fuzhou 8th High school.

Week19    8/7-14/7

1.Made 16s PCR of Staphylococcus epidermidis.[Result]
2.Did the lethal experiment of PLA against Staphylococcus epidermidis.
3.Knocked out Kana resistance of TnaA-/- deletion strain and hns-/- strain.
4.Amplified atoD and atoA [Result]
5.InterLab

Week21    15/7-21/7

1.Did the lethal experiment of PLA against Staphylococcus epidermidis.
2.Obtained the MazF toxin protein gene fragment from E.coli BW25113
3.The MazF and PEA32 backbones were connection by Gibsion. [Result]
4.Obtained lacI fragment by PCR. [Result]
5.InterLab

Week22    22/7-28/7

1.Attended the iGEM Eurasian Meetup 2018
2.Verified the function of MazF with different concentrations of arabinose concentration.[Result]
3.Did the T4 ligation for lacI and the circuit we synthesized.
4.Added different concentrations of PLA to samples and Measured OD value.
5.Tried to obtain eGFP fragment by PCR.
6.Uploaded the data of InterLab.

Week23    29/7-04/8

1.Making a standard curve of PLA.
2.Failed to constructed the circuit of hns-syn-eGFP, we decided to replace the eGFP with the GFP.
3.Change the backbone of the plasmid hns-syn from PUC57 into pRB1S.
4.Add restriction sites for atoD, atoA by PCR.
5.Did the lethal experiment of PLA against Staphylococcus epidermidis to provide data for our model.

Week24    5/8-11/8

1.Failed to enzyme cutting pRB1s, PLA and mazF with Kpn1 and Dra3.
2.Failed to construct pRB1s-atoD-atoA by Gibson assembly.
3.Obtained GFP fragment by PCR.
4.Tried to construct pRB1s-hns-syn-GFP.
5.Obtained ARO10, par and TyrB fragment by PCR. [Result]
6. Construct ARO10, par, TyrB and pSB1C3 by Gibson assembly.

Week25    12/8-18/8

1.Construct pSB1C3-atoD and pSB1C3-atoA by T4 connection.
2.Communicate with XMU-China.
3.The lethal experiment of mazf against Staphylococcus epidermidis.
4.Site-directed mutation.

Week26    19/8-25/8

1.The lethal experiment of mazf against Staphylococcus epidermidis.
2.T4 ligation for GFP and PLA.
3.Measure the production of 2-PE and PLA

Week27    26/8-01/9

Attended 5th ccic in Shanghai,China.

Week28    2/9-8/9

1.Got the fragment of adhE2.
2.Cultured our engineered bacteria in 0M, 0.05M, and 0.3M at 25℃ and 37℃.

Week29    9/9-15/9

1.Construct pRB1s-atoD-atoA-adhE2 and composite part. [Result]
2.T4 ligation for salt-temperature circuit and pSB1C3.
3.Line cultured the hns-syn on the different salt concentration of plates.

Week30    16/9-22/9

1.Observed the GFP level under fluorescence microscope.[Result]
2.verify whether circuit is successfully constructed.
3.Construct improve part.

Week31    23/9-29/9

Constructed araC+pBAD+tetR+GFP+PLtetO1+RFP part. [Result]

Week29    30/9-07/10

1.Submitted our parts
2.Made wiki
3.Verified circuit function

Lab Endding!
See our Lab book and protocol here!