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Team:Copenhagen/Results
From 2011.igem.org
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| + | <font color="#ffffff" face="constantia" size="8" align="left" > | ||
| + | <b><center>Results</center></b></font> | ||
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| - | <b><center> | + | <b><center>A2 the Igem Way</center></b><br><br> |
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| - | + | We mutated the CYP79A2 to remove a restrictionsite. Afterwards we inserted the CYP in the shipping plasmid. We confirmed both the mutation and the insertion by sequencing. | |
| + | We tried to insert the CYP79A2 in the ekspressionvector BBa_J04500 and although it looked like the gene was inserted when we got it sequenced it wasn't a perfect fit. We picked the best fitting and expressed them in BL21 cells. A SDS gel didn't show any increase in protein amount for the band-size corresponding to the CYP. A membrane preperation induced with CYPs substrate phenylalanine showed no production of oxime. | ||
| + | Our original A2 template have been reported to work (Ref to Kenneth article) and we therefore suspect the mutations and/or the prefix suffix to have influenced the activity. To explore this, we plan to analyse the device BBa_ K527001 - the cypa2 inserted in an expressionvector by the DTU_Denmark2 user assembly method which doesn't contain prefix/suffix but the mutation</p> | ||
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| - | <b><center> | + | <b><center>Killing fungus</center></b><br><br> |
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Revision as of 16:37, 17 September 2011
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We mutated the CYP79A2 to remove a restrictionsite. Afterwards we inserted the CYP in the shipping plasmid. We confirmed both the mutation and the insertion by sequencing. We tried to insert the CYP79A2 in the ekspressionvector BBa_J04500 and although it looked like the gene was inserted when we got it sequenced it wasn't a perfect fit. We picked the best fitting and expressed them in BL21 cells. A SDS gel didn't show any increase in protein amount for the band-size corresponding to the CYP. A membrane preperation induced with CYPs substrate phenylalanine showed no production of oxime. Our original A2 template have been reported to work (Ref to Kenneth article) and we therefore suspect the mutations and/or the prefix suffix to have influenced the activity. To explore this, we plan to analyse the device BBa_ K527001 - the cypa2 inserted in an expressionvector by the DTU_Denmark2 user assembly method which doesn't contain prefix/suffix but the mutation |
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