Part:BBa_K1061006

Bax mutant form

       Hbax, a member of Bcl-2 family, is an apoptosis-related protein. It integrates into the 

mitochondrial outer membrane, releases cytochrome C and leads to apoptosis.Hbax184a is a mutant of hbax that constantly integrates into mitochondrial outer membrane. We have tried to use them as suicide genes, but they can not induce apoptosis in most cancer cell lines, probably because the pathway that they involve has been blocked in most cancer cells. However, we find that the protein can induce “apoptosis” in the most broadly used eukaryotic chassis, yeast. So it will be very potential to use the gene in some circuit that need to kill the yeast in proper condition.

Calibration

    This is the pre-experment that we done.We inserted the hbax mutant form under the control of 
Gal promoter, then put it under the induction of the galatose. From left to right, the induction 

concentration is 0, 1%, and 2%. We can see a significant decrease of yeast concentration when put it in a high level of induction(2%)

pre-gradient experiment that we done.

quantutative analysis

  We have count the cell density of the yeast in the tude under 3 concentration, for the picture

below,top to down:0,1%,2%, it should be some treashold for the indution of apoptosis, so the gene behave somehow like a switch.

pre-gradient experiment that we have done data.

the data reveal in a picture form.