Difference between revisions of "Part:BBa K1061006"
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protein can induce “apoptosis” in the most broadly used eukaryotic chassis, yeast. So it will | 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. | be very potential to use the gene in some circuit that need to kill the yeast in proper condition. | ||
| + | We submitted as a improvement of already exist hbax biobrick. The link of original biobrick is | ||
| + | here:https://parts.igem.org/Part:BBa_K364202 | ||
| + | |||
====Calibration==== | ====Calibration==== | ||
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[[Image:Hbax_data.jpg|thumb|center|492×294|the data reveal in a picture form.]] | [[Image:Hbax_data.jpg|thumb|center|492×294|the data reveal in a picture form.]] | ||
| + | And, this year , SCUT also help us to characterize the part, by comparing its growth-inhibiting effect with the original hbax that we improve, we can see from the picture that the inhibition level of hbax mutant(label as hbaxs)is significantly higher than the hbax already exist.And they can both inhibit the growth comparing to the control. However, since 2% galactose is a very high indution level, it turns out that the highest expression level already reach at this point, so there are almost no difference between 2% and 5%. For more acurate gradient data, we need to set the gradient between 1% and 2% | ||
| + | [[Image:Hbax_and_hbax_mutant_comparing.jpg|thumb|center|722×412|the growth curve.]] | ||
Revision as of 04:39, 26 September 2013
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. We submitted as a improvement of already exist hbax biobrick. The link of original biobrick is here:https://parts.igem.org/Part:BBa_K364202
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%)
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.
And, this year , SCUT also help us to characterize the part, by comparing its growth-inhibiting effect with the original hbax that we improve, we can see from the picture that the inhibition level of hbax mutant(label as hbaxs)is significantly higher than the hbax already exist.And they can both inhibit the growth comparing to the control. However, since 2% galactose is a very high indution level, it turns out that the highest expression level already reach at this point, so there are almost no difference between 2% and 5%. For more acurate gradient data, we need to set the gradient between 1% and 2%
