Part:BBa_K1810002
] Melittin Optimized for E. coli with Mcherry
Melittin is a antimicrobial peptide found in bee venom that works to cause cell lysis by forming a pore in the membrane of harmful bacteria. While the 2014 Stony Brook iGEM team was able to successful clone the melittin gene into E. coli, expression was inconsistent. The 2015 Stony Brook iGEM team worked to correct this by optimizing the codons for an E. coli cell. Both the optimized and regular versions of melittin were fused with a mCherry tag in order to quantify differences in expression. With the use of a fluorescence plate reader, a near 100 fold increase in expression for the optimized melittin construct was observed. The untaged version of optimized melittin can be found in the following biobrick: BBa_K1810003
This graph shows the relative fluorescence levels of optimized melittin vs. regular melittin as expressed in E. coli as a function of arabinose concentration. As the melittin sequences were fused with a mCherry fluorescence tag, their level of translation could be observed with a plate reader. As the graph depicts, the optimization of melittin resulted in an approximately 100 fold increase in melittin protein production relative to that of unoptimized melittin. Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 65
Illegal XhoI site found at 947 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 193
Illegal AgeI site found at 185 - 1000COMPATIBLE WITH RFC[1000]
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