Team:SDU-Denmark/project-t

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(Hyperflagellation)
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== Hyperflagellation ==
== Hyperflagellation ==
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=== Background: ===<br>
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=== Background === <br>
To maximize the microflow system's effectivity, we want to increase the force each single bacterium can generate. Since the main motor for the flow is the flagellum we will have to modify this factor for increasing the force. Flagella in E.Coli rotate at a maximum speed around 6000 rpm, which can not easily be exceeded. So if we wanted to increse the generated force we would have to opt for more flagella on the surface of our bacteria, instead of faster flagella. This is called hyperflagellation, which is a process that is not all too well studied in normally-flagellated coli, so we will have to test it out.<br>
To maximize the microflow system's effectivity, we want to increase the force each single bacterium can generate. Since the main motor for the flow is the flagellum we will have to modify this factor for increasing the force. Flagella in E.Coli rotate at a maximum speed around 6000 rpm, which can not easily be exceeded. So if we wanted to increse the generated force we would have to opt for more flagella on the surface of our bacteria, instead of faster flagella. This is called hyperflagellation, which is a process that is not all too well studied in normally-flagellated coli, so we will have to test it out.<br>
The way we are hoping to achieve the hyper flagellation is bz upregulating the FlhD,C operon. This operon is the master regulator of flagella expression, which is controlled by three classes of regulons. The products of FlhD,C regulate the transcription of the class II factors, which in turn again regulate class III, FliC, the major subunit in flagella. Since FlhD,C is sitting on top of the regulating cascade we want to overexpress it, so that we will get an increase in flagellar count.<br>
The way we are hoping to achieve the hyper flagellation is bz upregulating the FlhD,C operon. This operon is the master regulator of flagella expression, which is controlled by three classes of regulons. The products of FlhD,C regulate the transcription of the class II factors, which in turn again regulate class III, FliC, the major subunit in flagella. Since FlhD,C is sitting on top of the regulating cascade we want to overexpress it, so that we will get an increase in flagellar count.<br>
The way we are going to achieve that is by isolating the operon from an E.Coli and inserting the coding sequence into a biobrick where we can control both the ribosome binding site and the type of promoter. We are going to use a constitutive promoter, so that flagella will be constantly expressed. The effects of this on other areas of the organism like the cell cycle are not entirely clear, but we will first try to overxpress the operon and then analyse the effect on the cells behavior.<br><br>
The way we are going to achieve that is by isolating the operon from an E.Coli and inserting the coding sequence into a biobrick where we can control both the ribosome binding site and the type of promoter. We are going to use a constitutive promoter, so that flagella will be constantly expressed. The effects of this on other areas of the organism like the cell cycle are not entirely clear, but we will first try to overxpress the operon and then analyse the effect on the cells behavior.<br><br>
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=== Biobrick design: ===<br>
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=== Biobrick design === <br>
FlhD,C coding sequence: BBa_K343000 (Sandboxed)
FlhD,C coding sequence: BBa_K343000 (Sandboxed)

Revision as of 17:56, 3 July 2010