Expression of Botulinum Neurotoxins A in Saccharomyces cerevisiae and Its Functional Research

Abstract:

Botulinum toxin（BoNT） is a neurotoxin produced by Clostridium botulinum，which is one of the most powerful toxin. To study the activity and function of BoNT/A in yeast，BoNT/A LC in yeast is expressed，observing the yeast growth and BoNT/A protein expressing level by western blot. Expression of LC of BoNT cerotype A caused a growth defect in yeast cells，and the toxicity is due to the hydrolytic activity. It is because catalytically inactive BoNT/A LC does not cause a growth defect in yeast cells. Overexpression of a yeast SNAP-25 homologue gene，SEC9，could rescue the growth defect in BoNT/A LC expressing cells. The results demonstrate that，in BoNT/A LC expressing yeast cells，the hydrolytic activity can be monitored through their growth. Thus，such cells can be used to analyze BoNT/A.