Difference between revisions of "Team:SDSZ China/Project"
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<p class="title">1.o</p> | <p class="title">1.o</p> | ||
<br/> | <br/> | ||
| − | <p></p> | + | <p>Principles： |
| + | 结构图 | ||
| + | PBP are ubiquitous bacterial enzymes involved in cell wall biosynthesis. | ||
| + | Penicillin-binding protein 5 (PBP 5) of Escherichia coli hydrolyzes the terminal D-Ala-D-Ala peptide bond of the stem precursor peptides of the cell wall peptidoglycan.(Investigation of the Mechanism of the Cell Wall DD-Carboxypeptidase Reaction of Penicillin-Binding Protein 5 of Escherichia coli by Quantum Mechanics/Molecular Mechanics Calculations | ||
| + | Qicun Shi, Samy O. Meroueh, Jed F. Fisher, and Shahriar Mobashery) | ||
| + | |||
| + | |||
| + | We designed a part that generates Penicillin Binding Protein 5-Green Fluorescence Protein (PBP5-GFP), and bind penicillin to the coated wells on the ELISA PLATE. The protein is added to the sample, where the penicillin in the sample compete for PBP5-GFP with those on the plate, and proteins unbound with penicillin stays on the plane. So we measure the fluorescence generated by GFP in the liquid, which indicates the content of the protein PBP5. With more penicillin present, more PBP5 binds to them, and greater fluorescence intensity will be measured. (See protein purification protocol) | ||
| + | |||
| + | Proof of concept: | ||
| + | 图：蛋白质电泳＋解析 | ||
| + | |||
| + | |||
| + | Results: | ||
| + | 图：用青霉素溶液做的结果图＋解析 | ||
| + | See protocol | ||
| + | |||
| + | |||
| + | Demonstration: | ||
| + | 图：用牛奶测得结果图＋解析 | ||
| + | See Protocol | ||
| + | |||
| + | |||
| + | Optimization: | ||
| + | 改分光光度计</p> | ||
</div> | </div> | ||
</div> | </div> | ||
Revision as of 09:40, 14 October 2016
1.o
Principles： 结构图 PBP are ubiquitous bacterial enzymes involved in cell wall biosynthesis. Penicillin-binding protein 5 (PBP 5) of Escherichia coli hydrolyzes the terminal D-Ala-D-Ala peptide bond of the stem precursor peptides of the cell wall peptidoglycan.(Investigation of the Mechanism of the Cell Wall DD-Carboxypeptidase Reaction of Penicillin-Binding Protein 5 of Escherichia coli by Quantum Mechanics/Molecular Mechanics Calculations Qicun Shi, Samy O. Meroueh, Jed F. Fisher, and Shahriar Mobashery) We designed a part that generates Penicillin Binding Protein 5-Green Fluorescence Protein (PBP5-GFP), and bind penicillin to the coated wells on the ELISA PLATE. The protein is added to the sample, where the penicillin in the sample compete for PBP5-GFP with those on the plate, and proteins unbound with penicillin stays on the plane. So we measure the fluorescence generated by GFP in the liquid, which indicates the content of the protein PBP5. With more penicillin present, more PBP5 binds to them, and greater fluorescence intensity will be measured. (See protein purification protocol) Proof of concept: 图：蛋白质电泳＋解析 Results: 图：用青霉素溶液做的结果图＋解析 See protocol Demonstration: 图：用牛奶测得结果图＋解析 See Protocol Optimization: 改分光光度计
2.0
3.0
