Team:Jiangnan China/Notebook

• Project
  • Inspiration and Description
  • Proof of Concept
  • Implementation
• Dry Lab
  • Model
• Wet Lab
  • Parts
  • Engineering Success
  • Results
• Human Practices
  • Integrated Human Practices
  • Collaboration
  • Partnership
• Science Communication
• Team
  • Team members
  • Attribution
  • Notebook
• Future
  • Contribution
  • Prospect

• MAY
• JUN
• JUL
• AUG
• SEP
• OCT

1. Week1 MAY 1-MAY 2

Codon optimization of green fluorescent protein (GFP) gene sequence was carried out.

2. Week2 MAY 3-MAY 9

Promoter Ppgki was used to express the gene gfp. The recombinant plasmid with the gene gfp was integrated into Starmerella Bombicola. Fluorescence intensity of the recombinant strains were measured using laser scanning confocal microscope. According to the results, it was verified that GFP could be successfully expressed in S. Bombicola, which could be used as a reporter gene for promoter screening.

3. Week3 MAY 10-MAY 16

After sequence analysis of S. Bombicola whole genome, 13 promoter sequences were obtained by PCR using specific designed primers.

4. Week4 MAY 17-MAY 23

The fusion structures “promoter-GFP-terminator” (Ptef1, Ptef2, Pgpd, Peno, Ptpi and Ptdh1 ) were assembled by fusion PCR and inserted into plasmid pPHP to obtain the recombinant plasmids, which were respectively linearized and then electroporated to the wild-type S. Bombicola to obtain the recombinant strains.

5. Week5 MAY 24-MAY 30

Pfao, Ppgi, Padh1, Pcyp52m1, Pmob and Padh1 were fused with GFP by fusion PCR and inserted into the starting plasmid pPHP to obtain the recombinant plasmids, which were respectively linearized and then electroporated to the wild-type S. Bombicola to obtain the recombinant strains.

6. Week6 MAY 31

Promoters of UGTA1 and UGTB were fused with the gene gfp by fusion PCR and inserted into the starting vector pPHP to obtain the recombinant plasmids, which were linearized and then electroporated to the wild-type S. Bombicola to obtain the recombinant strains, respectively.