Difference between revisions of "Part:BBa K5115070"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K5115070 short</partinfo> | <partinfo>BBa_K5115070 short</partinfo> | ||
| − | <html><img style="float:right;width:128px" src="https://static.igem.wiki/teams/5115/czh/mineral-logo.svg" alt="contributed by Fudan iGEM | + | <html><img style="float:right;width:128px" src="https://static.igem.wiki/teams/5115/czh/mineral-logo.svg" alt="contributed by Fudan iGEM 2024"></html> |
__TOC__ | __TOC__ | ||
===Introduction=== | ===Introduction=== | ||
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bound AAA+ protease, and its protease activity toward LpxC is regulated by | bound AAA+ protease, and its protease activity toward LpxC is regulated by | ||
essential membrane proteins LapB and YejM. YejM can compete with FtsH for LapB to inhibit LpxC | essential membrane proteins LapB and YejM. YejM can compete with FtsH for LapB to inhibit LpxC | ||
| − | degradation, thus guaranteed the expression level of LPS | + | degradation, thus guaranteed the expression level of LPS <ref>Shu, S., & Mi, W. (2022). Regulatory mechanisms of lipopolysaccharide synthesis in Escherichia coli. Nature Communications, 13(1), 4576.</ref>. LPS is a basic component of ''E.coli's'' outer membrane. Sufficient expressing of LPS can help with the bacteria's survival. So, as a result, the YejM can indirectly improve ''E.coli's'' viability. |
| + | |||
| + | {| | ||
| + | | <html><img style="width:400px" src="https://static.igem.wiki/teams/5115/part-collection/parts-anti-phage.png" alt="contributed by Fudan iGEM 2024"></html> | ||
| + | |- | ||
| + | | '''Figure 1. The structure and function of YejM. | ||
| + | ''' | ||
| + | |||
| + | |} | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
Our MINERAL is designed to be put into half-open water. To ensure that it can function stably, we choose to use YejM to raise ''E.coli'''s viability. | Our MINERAL is designed to be put into half-open water. To ensure that it can function stably, we choose to use YejM to raise ''E.coli'''s viability. | ||
===Characterization=== | ===Characterization=== | ||
| + | We infected ''E. coli'' TG1 with kanamycin-resistant M13KO7 phage and plated a 200-fold dilution on kanamycin plates. This allowed us to assess the phage infection efficiency by counting the number of colonies. According to our results, it's clear that TG1-YejM significantly exhibits better growth than TG1-stayGold. YejM enables ''E. coli'' to thrive in the presence of phages, demonstrating its effectiveness in providing anti-phage capability. | ||
| + | |||
| + | {| | ||
| + | | <html><img style="width:400px" src="https://static.igem.wiki/teams/5115/txj/10-5.jpg" alt="contributed by Fudan iGEM 2024"></html> | ||
| + | |- | ||
| + | | '''Figure 2. Phage Colony Formation, with and without YejM expression. | ||
| + | The ''E. coli'' TG1 carrying either stayGold fluorescent protein and YejM under the J23107 promoter were infected with M13KO7 phage at different MOIs. Colonies were counted after incubating at 37°C for 16 hours, on selection LB plates. "*" indicates a p-value less than 0.01. Under any MOI condition, the colony count of ''E. coli'' TG1 carrying YejM was significantly lower than that of stayGold, indicating that YejM expression confers resistance to phage infection. | ||
| + | ''' | ||
| + | |} | ||
| + | ===Sequence and Features=== | ||
<!-- --> | <!-- --> | ||
<span class='h3bb'>Sequence and Features</span> | <span class='h3bb'>Sequence and Features</span> | ||
Latest revision as of 12:25, 2 October 2024
YejM
Introduction
YejM is an anti-adaptor protein that can regulate the synthesis of the LPS. In E.coli, LPS levels are under feedback control, achieved by FtsH-mediated degradation of LpxC. FtsH is a membrane- bound AAA+ protease, and its protease activity toward LpxC is regulated by essential membrane proteins LapB and YejM. YejM can compete with FtsH for LapB to inhibit LpxC degradation, thus guaranteed the expression level of LPS [1]. LPS is a basic component of E.coli's outer membrane. Sufficient expressing of LPS can help with the bacteria's survival. So, as a result, the YejM can indirectly improve E.coli's viability.
|
| Figure 1. The structure and function of YejM.
|
Usage and Biology
Our MINERAL is designed to be put into half-open water. To ensure that it can function stably, we choose to use YejM to raise E.coli's viability.
Characterization
We infected E. coli TG1 with kanamycin-resistant M13KO7 phage and plated a 200-fold dilution on kanamycin plates. This allowed us to assess the phage infection efficiency by counting the number of colonies. According to our results, it's clear that TG1-YejM significantly exhibits better growth than TG1-stayGold. YejM enables E. coli to thrive in the presence of phages, demonstrating its effectiveness in providing anti-phage capability.
|
| Figure 2. Phage Colony Formation, with and without YejM expression.
The E. coli TG1 carrying either stayGold fluorescent protein and YejM under the J23107 promoter were infected with M13KO7 phage at different MOIs. Colonies were counted after incubating at 37°C for 16 hours, on selection LB plates. "*" indicates a p-value less than 0.01. Under any MOI condition, the colony count of E. coli TG1 carrying YejM was significantly lower than that of stayGold, indicating that YejM expression confers resistance to phage infection. |
Sequence and Features
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1440
- 1000COMPATIBLE WITH RFC[1000]
References
- ↑ Shu, S., & Mi, W. (2022). Regulatory mechanisms of lipopolysaccharide synthesis in Escherichia coli. Nature Communications, 13(1), 4576.