Team:SUSTech Shenzhen/Notebook


Notebook of Project

About Notebook



Quorum sensing consultation (target Vibrio cholera)

Salt promoter consultation (chloride induced promoter)

pH promoter consultation (induced at pH5.5)

Design of the circuit

Colonic acid assay methodology

Literature research about antimicrobial peptides(AMPs)

Purchase chemicals and gene

Plasmid design of Lactobacillus

Purification of human lactoferrin(hLF)


Antibacterial test of hLF on V.cholera and E. coli Nissle 1917

CAI-1 quorum sensing modeling

Culture and preservation of Lactobacillus

Antibacterial Test(hLF, 7475 and kanamycin on V.cholera)

Make competent cell of Nissle 1917, Lactobacillus

Plasmid extraction of pBbE6k and pTRKH3-slpGFP

Transformation of pBbE6k into Nissle 1917

Plasmid construction of RcsB – pBbE6k(digestion and ligation)

Purification of hLF and 7475

Transformation of pBbE6k into E. coli Top10


Transformation of RcsB – pBbE6k and control into E. coli DH5α and Top10

Colony PCR and sequencing of Top10+pBbE6k, pBbE6k-RcsB

Purification of hLF

Sequencing of RcsB – pBbE6k and control into Top10

Plasmid extraction of pBbE6k

Transformation of pBbE6k into E. coli BL21 and E. coli Acella

Antibacterial Test


IPTG induced RcsB expression, electrophoresis, western blot in Acella and BL21

IPTG induced expression of RFP in Nissle 1917

Transformation of pBbE6k-RcsB into Nissle 1917

Staining and observation of BL21 and Acella with RcsB expression

Design of experiment of hLF


Transformation of RcsB into Nissle 1917

the Lactoferrin test

Verify the chloramphenicol function



Western blot of cholera toxin detection in Vibric cholera BNCC 232030


Five schools exchange meeting

Cooperation negotiation with SZU-China

Counseling from Feiru Biotech

Counseling from doctors at the First Affiliated Hospital of Xi’an Jiaotong University


Expression and verification of RcsB in Nissle using IPTG

Construction of plasmids (CAI-1 detector, lac-ara-induced(EsaR), Lactobacillus gadR)

Counseling from doctors at the Shenzhen Second People's Hospital

Deep Blue Charity Sale

Communication with Central South University (CSU)

Counseling from scientist

Spring buds lecture

China iGEM Online Meetup


Learn and practice molecular cloning methods and other methods, including PCR, Gibson Assembly, plasmid transformation, Western Blot, HPLC etc

Communication with the National University of Singapore (NUS)

Spring buds lecture

"Cloud West" online volunteering teaching project


Preliminary purification of colanic acid in Nissle

Transformation EsaR plasmid into BL21


Further literature research about UHPLC assay

Preparation of competent cell Acella and DH5α

Transformation of CAI-I and EsaR plasmid into Acella and gadR plasmid into DH5α

Preservation of transformed bacteria Acella and DH5α


Optimize the condition for UHPLC assay (different mobile phases)

SUSTech Hospital Exchange

Gastrointestinal meeting


Set up four standard curves of monosaccharides using UHPLC assay

Preservation of transformed bacteria DH5α with PKRTH3-ermB

PCR of arabinose promoter

PCR of vector(lac-ara-induced(EsaR))



Optimize the condition for UHPLC assay (different flow rate, time constant, reference standards)

DNA electrophoresis for fragment(arabinose promoter) and vector Gibson

Transformation of the product of Gibson

Preservation of transformed bacteria


Optimize the condition for UHPLC assay (different gradient methods)

Western blot of PhlF in BL21

Spring buds lecture


Modification to get a better separation under more detailed conditions for UHPLC assay

PCR of the colony from Gibson


Western blot of EsaR, CqsS, LuxU, LuxO in Acella