Team:Washington/Magnetosomes/Results

From 2011.igem.org

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Using standard synthetic biology protocols and the vectors we created in our Gibson Assembly Toolkit, our team was able to create a '''"Magnetosome Toolkit"''' consisting of the most basic parts required for magnetosome formation. Providing this toolkit allows future iGem teams to manipulate and understand magnetosome formation to one day create magnets in various types of bacteria.  
Using standard synthetic biology protocols and the vectors we created in our Gibson Assembly Toolkit, our team was able to create a '''"Magnetosome Toolkit"''' consisting of the most basic parts required for magnetosome formation. Providing this toolkit allows future iGem teams to manipulate and understand magnetosome formation to one day create magnets in various types of bacteria.  
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===What' in the toolkit and How we assembled it===
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===What' in the toolkit and How We Assembled It===
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| 1002
| 1002
|-
|-
-
  |}
+
  |}.
Using standard PCR protocols, these genes were extracted and.....
Using standard PCR protocols, these genes were extracted and.....
These genes were visualized on gels and sequence confirmed; therefore, we can be sure that these are some of the many genes required for proper magnetosome formation.
These genes were visualized on gels and sequence confirmed; therefore, we can be sure that these are some of the many genes required for proper magnetosome formation.
 +
As previously noted, magnetosome formation within the host-organism, ''Magnetospirillium magneticum'', strain AMB-1, is a highly regulated step-wise process. As shown in (link: figure #), genes encode for an invagination in the inner membrane, there are genes which help align the magnetosomes into their characteristics chains, and there are genes which regulate the biomineralization of magnetic particles. Our team chose to focus on genes specifically related to magnetosome scaffolding/alignment, since it has many practical uses in synthetic biology (??? maybe we could make a link to the alkenes future if they wanted to use mamI....)
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However, in order to make sure that gene-activity had not be lost, we created mam gene-sfGFP fusions, transformed them into E.coli, and imaged them with microscopy. The two most notable constructs were mamK-sfGFP and mamI-sfGFP
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Our genes of interest were mamK and mamI as they have functions related to localization of the magnetosome. Specifically, mamK is a bacterial actin-like cytoskeleton protein required for proper alignment of the magnetosomes in a chain. mamK is also shown to localize the mamI, which is loss inhibits membrane formation.
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For more information, please refer to the table below:
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(--> we're going to put pictures here....<--)
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{| class="wikitable"
 +
|-
 +
! Gene
 +
! AMB Number
 +
! Cluster Membership
 +
! Member of 28 genes list? (specific*/related**)
 +
! Function Summary (Vesicle chain formation, and/or biomineralization)
 +
! Gene Function
 +
|-
 +
| mamHI
 +
| 1541
-
-essential gene
+
| mamE
-
a picture of gel, notable constructs, process diagram, and cool results please.
+
| 2172
 +
|-
 +
| mamJ
 +
| 1538
 +
|-
 +
| mamKL
 +
| 1336
 +
|-
 +
| mamMN
 +
| 2323
 +
|-
 +
| mamO
 +
| 1914
 +
|-
 +
| mamPA
 +
| 1493
 +
|-
 +
| mamQRB
 +
| 2029
 +
|- 
 +
| mamSTU
 +
| 2030
 +
|-
 +
| mamV
 +
| 1002
 +
|-
 +
|}.

Revision as of 23:11, 18 September 2011


About the Magnetosome Toolkit:

Washington Methode image.jpg

Using standard synthetic biology protocols and the vectors we created in our Gibson Assembly Toolkit, our team was able to create a "Magnetosome Toolkit" consisting of the most basic parts required for magnetosome formation. Providing this toolkit allows future iGem teams to manipulate and understand magnetosome formation to one day create magnets in various types of bacteria.

What' in the toolkit and How We Assembled It

After piecing together the 16 kb genome of the mamAB gene cluster within the magnetosome island (MAI), we extracted out the genes in the following group:

Gene groups Length (bp)
mamHI 1541
mamE 2172
mamJ 1538
mamKL 1336
mamMN 2323
mamO 1914
mamPA 1493
mamQRB 2029
mamSTU 2030
mamV 1002
.

Using standard PCR protocols, these genes were extracted and..... These genes were visualized on gels and sequence confirmed; therefore, we can be sure that these are some of the many genes required for proper magnetosome formation.

As previously noted, magnetosome formation within the host-organism, Magnetospirillium magneticum, strain AMB-1, is a highly regulated step-wise process. As shown in (link: figure #), genes encode for an invagination in the inner membrane, there are genes which help align the magnetosomes into their characteristics chains, and there are genes which regulate the biomineralization of magnetic particles. Our team chose to focus on genes specifically related to magnetosome scaffolding/alignment, since it has many practical uses in synthetic biology (??? maybe we could make a link to the alkenes future if they wanted to use mamI....)


Our genes of interest were mamK and mamI as they have functions related to localization of the magnetosome. Specifically, mamK is a bacterial actin-like cytoskeleton protein required for proper alignment of the magnetosomes in a chain. mamK is also shown to localize the mamI, which is loss inhibits membrane formation. For more information, please refer to the table below:

Gene AMB Number Cluster Membership Member of 28 genes list? (specific*/related**) Function Summary (Vesicle chain formation, and/or biomineralization) Gene Function
mamHI 1541 mamE 2172
mamJ 1538
mamKL 1336
mamMN 2323
mamO 1914
mamPA 1493
mamQRB 2029
mamSTU 2030
mamV 1002
.