Team:UMaryland/Notebook
Notebook
Week 1
Miraculin:
ligated the pBAD promoter in PSB1C3 with the miraculin gene in PSB1C3 using 3A assembly
plate had colonies
Week 2
Miraculin:
pBAD +miraculin construct was moved back into psb1C3 backbone and sent for sequencing
Designing gBlocks:
Designed gblocks for epsilon-cyclase (from arabidopsis), pyocin and Hok/Sok
Week3
Miraculin:
sequence confirmed through sequencing
failed to extract using French press, FPLC and SDS-Page
could be due to high arabinose induction (OD of 1)
Week 4
Miraculin:
induced 3 test cultures with 0%, 0.05%, 0.1%, 0.2% arabinose with no significant results
will perform procedure again using bl21 instead of Dh5a
Week 5
Miraculin
transformation in BL21 strain was successful
SDS page showed a band at roughly 25 kDa
Hok/Sok
inserted Hok/Sok gblock into PSB1C3 backbone using Gibson assembly
construct sent for sequencing
Interlab Study
transformed parts into dh5a: 3 Anderson promoters, GFP, and RFP
Week 6
Miraculin:
induced 3 test cultures with 0%, 0.05%, 0.1%, 0.2% arabinose with no significant results
will perform procedure again using bl21 instead of Dh5a
Week 13
Miraculin:
Week 14
Miraculin:
Week 15
Miraculin:
Week 16
Miraculin: