Difference between revisions of "Part:BBa K3888008:Experience"

 
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter
 
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how you used this part and how it worked out.
 
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===Applications of BBa_K3888008===
 
===Applications of BBa_K3888008===
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[[File:Deletion Plasmid construction.png]]
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===Figure 1: Illustration  of the general form of plasmid used to create marked knockout strains of R. palustris. ===
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The origin of replication (ori), origin of transfer (oriT) and sacB (blue) are part of the delivery plasmid. The green sequence is inserted into the multiple cloning site of the delivery plasmid. The left and right fragments (green) are sequences amplified from the R. palustris genome and used in overlap extension PCR to generate a recombinant product containing (a deletion and a restriction site）. The restriction site is used to introduce the resistance cassette (purple). This interrupts the gene even if no deletion is present, and allows selection of successful transformants. <br>
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According to this model, we constructed UppE knockout plasmid below.<br>
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[[ File:HupS Deletion Plasmid Map.png]]
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===Figure 2: Deletion plasmid of HupS constructed according to the method mentioned above.===
  
 
===User Reviews===
 
===User Reviews===
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Reference: Rey, Federico E., Yasuhiro Oda, and Caroline S. Harwood. "Regulation of uptake hydrogenase and effects of hydrogen utilization on gene expression in Rhodopseudomonas palustris." Journal of bacteriology 188.17 (2006): 6143-6152.
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Enter the review inofrmation here.
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Revision as of 19:45, 20 October 2021


Applications of BBa_K3888008

Deletion Plasmid construction.png

Figure 1: Illustration of the general form of plasmid used to create marked knockout strains of R. palustris.

The origin of replication (ori), origin of transfer (oriT) and sacB (blue) are part of the delivery plasmid. The green sequence is inserted into the multiple cloning site of the delivery plasmid. The left and right fragments (green) are sequences amplified from the R. palustris genome and used in overlap extension PCR to generate a recombinant product containing (a deletion and a restriction site）. The restriction site is used to introduce the resistance cassette (purple). This interrupts the gene even if no deletion is present, and allows selection of successful transformants.
According to this model, we constructed UppE knockout plasmid below.
HupS Deletion Plasmid Map.png

Figure 2: Deletion plasmid of HupS constructed according to the method mentioned above.

User Reviews

UNIQ0d1d186fd18af49b-partinfo-00000000-QINU
Reference: Rey, Federico E., Yasuhiro Oda, and Caroline S. Harwood. "Regulation of uptake hydrogenase and effects of hydrogen utilization on gene expression in Rhodopseudomonas palustris." Journal of bacteriology 188.17 (2006): 6143-6152. UNIQ0d1d186fd18af49b-partinfo-00000001-QINU