Established an IGEM team and conducted online meeting every weeks
Discussed and chose the overall direction of the project
Selected team leaders and organized subsequent division of labor based on the strengths of each member
Learn theoretical knowledge related to synthetic biology together
4.15 - 7.13
Conduct desk research to understand the current market demand and existing related pure natural polymer materials in the market
Based on the current resistance of people to antibiotics, it is proposed to introduce antimicrobial peptides into materials and endow them with antibacterial properties
Review literature to find suitable protein expression systems and appropriate antimicrobial peptides
7.14
Went through the local laboratory’s safety protocol and iGEM official safety guideline
Constructed plasmids containing CP1 gene
Used sodium carbonate solution to steam the silk to wash away the silk gelatin protein
Dried the separated silk protein
7.15
Constructed plasmids containing CP1 gene
Used sodium carbonate solution to steam the silk to wash away the silk gelatin protein
Dried the separated silk protein
Degummed silk into the ternary solution stirring and dissolution for 3h after filtration
Dialysed the filtered solution
7.16
Imported plasmids into E.coil to construct recombinant expression strains
Unsuccessful in obtaining recombinant expression strains
Used sodium carbonate solution to steam the silk to wash away the silk gelatin protein
Dried the separated silk protein
Degummed silk into the ternary solution stirring and dissolution for 3h after filtration
Dialysed the filtered solution
7.17
Designed a cell-free expression system
Successfully expressed CP1 antimicrobial peptide
Used sodium carbonate solution to steam the silk to wash away the silk gelatin protein
Dried the separated silk protein
Degummed silk into the ternary solution stirring and dissolution for 3h after filtration
Dialysed the filtered solution
7.18
SDS-PAGE and WB blot were used to verify the protein expression level
Found that the amount of expression was too small, and the gap between the equivalent amount and the actual use was too large, so the antimicrobial peptide was abandoned
7.19
Reviewed the literature and started to use M2, an antimicrobial peptide that is toxic only after enzymatic cleavage.
Constructed a plasmid containing the M2 gene
Conducted field research and distributed questionnaires for interviews
7.20
Imported the plasmid containing the M2 gene into E. coli to construct a recombinant expression strain
Visited a number of fruit and vegetable supermarkets to conduct interviews
7.21
Added IPTG to induce the recombinant strain to express the antimicrobial peptide
Visited a number of fruit and vegetable supermarkets to conduct interviews
7.22
Lysed the bacteria
Purified by affinity chromatography
Used OD600 detection to check bacterial inhibitory activity
Soaked freshly cut apple lettuce with different concentrations of silk protein and other common preservatives
Controlled experiments were carried out
The weight loss of fruits and vegetables was measured daily
7.23
The weight loss of fruits and vegetables was measured daily
Did microbiological analysis to determine the growth of bacteria on fruits and vegetables
Interviewed professors from related industries
Made a promotion video
Drew the logo of the team
Promoted the team through bilibili, Xiaohongshu and other channels
7.24
The weight loss of fruits and vegetables was measured daily
Did microbiological analysis to determine the growth of bacteria on fruits and vegetables
Interviewed professors from related industries
Made a promotion video
Drew the logo of the team
Promoted the team through bilibili, Xiaohongshu and other channels
7.25
The weight loss of fruits and vegetables was measured daily
Did microbiological analysis to determine the growth of bacteria on fruits and vegetables