Part:BBa_M36087:Experience

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Applications of BBa_M36087

Winter Flounder Antifreeze Protein Actuator without Secretion Tag

We had yeast take up the Winter Flounder Antifreeze Protein (AFP) Actuator without the yeast secretion tag and we tested the function of the actuator under cold conditions.

Results:

1) Significant difference in colony number

The results suggest that the actuator was expressed because the colonies lived up to 24 hours after freezing in the -80 degree freezer. There was a significant difference between our positive and negative controls (our negative control also showed similar growth after freezing for the same amount of time). This is believed to have occurred because there was a leaky promoter because the yeast was exposed to extreme cold temperatures. However, the negative control yeast were put in the -80 degree for 24 hours and still grew up, which suggests that there was some AFP expression.

2) Bigger diameter of colonies that have the induced actuator

We also found that colonies that grew up after 24 hours of freezing had larger diameters, implying that they are either healthier cells or that there is more room for them to grow. However, we did not find a significant difference in number of cells, therefore it could suggest the former.

Status of Yeast Secretion Tag:

Either Minor Benefits or no Effect on AFP actuator function:

There was no significant difference in growth between colonies that have the yeast secretion tag and those that do not. It also seems that the yeast secretion tag did not hinder the AFP expression meaning that it either had beneficial or no effect. Further testing is needed to verify the role of the yeast secretion tag.

Future Steps:

1) Greater time intervals in the cold temperature (28 h, 34 h, 40h, 48h) 2) Negative control with glucose to repress the galactose sensor are also needed to see whether the actuator was fully functional

Conclusion:

However, our results suggest that the actuator functions to some degree.

Stanford Location:

-80 C Freezer

Box Label: BioE44 S14

Device Type: Actuator

Organism: Yeast

DNA 2.0 Gene#: 151275

Barcode Number:0133011386

Plasmid Name: pD1201

User Reviews

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