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Team:Groningen/Project/Motility
From 2013.igem.org
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<h2>Motility</h2> | <h2>Motility</h2> | ||
<p> | <p> | ||
| - | + | Bacterial movement is based on flagella (tail like structures) and utilizes a counter-clockwise (CWW) | |
| - | + | and clockwise (CW) motion. When the flagella turn CCW they gather in one area resulting in | |
| - | + | bacteria that move straight. When the flagella move CW they disperse all over the cell membrane, | |
| - | Bacterial movement based on flagella (tail like structures) and utilizes | + | resulting in the bacteria tumbling in random directions. When bacteria sense an |
| - | and CW | + | attractant the flagellar motion will turn CCW, when the concentration of the attractant reduces the flagella will turn CW. |
| - | bacteria that move | + | |
| - | resulting in the bacteria | + | |
| - | attractant | + | |
| - | + | ||
</p> | </p> | ||
| - | <h3>The | + | <h3>The principle</h3> |
<p> | <p> | ||
The receptor is displayed in the figure on the page below. The letters are all Che proteins, with this | The receptor is displayed in the figure on the page below. The letters are all Che proteins, with this | ||
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(by forming a CheY-p/CheC complex) that CheD is pulled off the receptor. CheD and CheC forms | (by forming a CheY-p/CheC complex) that CheD is pulled off the receptor. CheD and CheC forms | ||
also a complex which dephosphorylizes CheY-p into CheY thus resetting the receptor. | also a complex which dephosphorylizes CheY-p into CheY thus resetting the receptor. | ||
| - | <Br> | + | <Br><br> |
CheY is a mayor factor in spinning the flagella CCW. When <i>cheY</i> is absent, cells are significant less motile[ref]. Because the promoter of <i>des</i> is active at low temperatures (25 °C) we placed <i>cheY</i> under control of the promoter of <i>des</i> (figure 2). | CheY is a mayor factor in spinning the flagella CCW. When <i>cheY</i> is absent, cells are significant less motile[ref]. Because the promoter of <i>des</i> is active at low temperatures (25 °C) we placed <i>cheY</i> under control of the promoter of <i>des</i> (figure 2). | ||
<br> | <br> | ||
Revision as of 14:33, 8 September 2013
Heat Motility
In case of a low yield we want a targeted secretion only near our (that we want to coat with silk). In order to achieve this we want to have a bacillus that will move towards heat. If the implant is heated it will attract our silk secreting bacillus.
The cold sensor DesK
Use of DesK as an sensor for temperature. DesK is a membrane protein (a kinase), it is part of a cascade that has to maintain the membrane liquidity. It does this by phosphorylating DesR, this phosphorylated DesR activates a promotor (pdes) that expresses the des gene, changing the membrane in the process.
The promoter activity of des
(a)Pattern of Pdes-lacZ expression on a temperature downshift. B. subtilis AKP3 cells were grown at 37 °C to an optical density of 0.4 at 525 nm and then divided into two fractions. The first was transferred to 25 °C (●) and the second was kept at 37 °C (○). (b) Pattern of Pdes-lacZ expression in a des‾ background. B. subtilis AKP4 cells were grown at 37 °C to an optical density of 0.4 at 525 nm and then divided into two fractions. One fraction was transferred to 25 °C (●) while the other was kept at 37 °C (○). (c). Effect of exogenous fatty acids on Pdes-lacZ expression pattern. B. subtilis AKP4 cells were grown at 37 °C to an optical density of 0.4 at 525 nm and then divided into two fractions. Each fraction was supplemented with palmitic (●) or oleic acid (■) and growth was continued at 25 °C. (d) Effect of desKR disruption on Pdes-lacZ expression. B. subtilis AKP21 cells were grown at 37 °C to an optical density of 0.4 at 525 nm and then divided into two fractions. One of the fractions was transferred to 25 °C (●) and the other one was kept at 37 °C (○). Optical density at 525 nm (inserts) and β-galactosidase specific activity were determined at the indicated times (a, b, c, or d).
Motility
Bacterial movement is based on flagella (tail like structures) and utilizes a counter-clockwise (CWW) and clockwise (CW) motion. When the flagella turn CCW they gather in one area resulting in bacteria that move straight. When the flagella move CW they disperse all over the cell membrane, resulting in the bacteria tumbling in random directions. When bacteria sense an attractant the flagellar motion will turn CCW, when the concentration of the attractant reduces the flagella will turn CW.
The principle
The receptor is displayed in the figure on the page below. The letters are all Che proteins, with this
cascade of proteins motility is coordinated. When a attractant is bound to the receptor, CheA
phosphorylizes CheY into CheY-P. CheY-P causes the CCW motility in the flagella, it also causes
(by forming a CheY-p/CheC complex) that CheD is pulled off the receptor. CheD and CheC forms
also a complex which dephosphorylizes CheY-p into CheY thus resetting the receptor.
CheY is a mayor factor in spinning the flagella CCW. When cheY is absent, cells are significant less motile[ref]. Because the promoter of des is active at low temperatures (25 °C) we placed cheY under control of the promoter of des (figure 2).
References
Mariana Martin and Diego de Mendoza , Regulation of Bacillus subtilis DesK thermosensor by lipids, Biochemical Journal (2013), Vol 451 No 2, pp. 269–275
Christopher V. Rao, George D. Glekas and George W. Ordal, The three adaptation systems of Bacillus subtilis chemotaxis, Trends in Biology (2008), Vol. 16 No 10, pp. 480-487.
Liam F. Garrity and George W. Ordal, Chemotaxis in Bacillus Subtilis: How bacteria monitor environmental signals, Pharmacology and Therepeutics (1995), Vol. 68 No.1, pp. 87-104.
