Team:UNIK Copenhagen/Results
Project Results
A few days after transformation we observed the moss protoplasts under a fluorescent microscope and saw YFP expression in moss protoplats for the antifreeze and the STS construct (fig. 4A and 4B). This confirms the that our transformation was a success and highly suggests that our genes of interest, antifreeze and STS, are expressed. It demonstrated that our construct design works and that moss can combine different DNA pieces with matching overhangs using homologous recombination.
Figure 4: Fluorescence microscopy pictures of P. patens transformed with our genetic constructs. A) A moss protoplast transformed with the antifreeze construct. B) A moss protoplast transformed with our STS construct. C) A moss protoplast transformed with a vector expressing YFP. A positive control. D) WT moss. A negative control. 1) Bright field picture. 2) Filter showing autoflouorescence (red) and YFP-expression (green). 3) Filter showing only YFP-expression (green).
Our transformed moss protoplasts were then moved to to PhyB-plates containing kanamycin (50 mg/ml) and were left to grow for a few weeks. One month after transformation we had ten growing clumps of transformed moss. Seven of those clumps of moss were expressing YFP (fig. 4). This suggests stable integration of our gene constructs.
Figure 5: A clump of transformed P. Patens showing YFP-expression, grown on kanamycin containing plates (50 mg/ml). A) Bright field picture. B) Filter showing autoflouorescence (red) and YFP-expression (green).
C) Filter showing only YFP-expression (green).