Part:BBa_K2052014
FimH site directed mutated with RPMrel
Usage and Biology
Some substrains of E.Coli have a structure called Type 1 pili which is expressed from the Fim gene system. At the end of the pili structure there is a protein called “FimH” which is the structure that allows them to bind to the mannose sugar that is found on the surfaces of eukaryotic cells. (Sauer et al., 2016).However, the substrain that was used in this project was BL21, a non-pathogenic laboratory strain. Deleted mannose binding and replacing it with RPMrel would provide tumor specific binding (Kelly et al., 2003).The CPIEDRPMC (RPMrel) peptide can bind to five colon cancer cell lines: HT29, CaCo-2, RKO, SW480, and DLD-1. Here on we have choosen CaCo-2 that is studied commonly in METU as our candidate to show targeted thearpy.
3D Structure of FimH together with RPMrel can be seen below as Harvard BioDesign 2015 submitted in part registry
ITU takımı bize RPMrelli bağlanabilen gif gönderecek. Onu tam bunun altına yerleştir.
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DNA Gel Analysis
. In the first 4 lanes the the gene is shown approximately 2100 base pairs and it is uncut. In the firts 4 lanes after the second ladder the gene is shown approximately 3300 base pairs and it is single cut with EcoRI. With the single cut it’s been shown that the insert is successfully in our vector.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1173
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1113
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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