Difference between revisions of "Part:BBa K346033: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.
 
  
 
===Applications of BBa_K346033===
 
===Applications of BBa_K346033===
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[[dsba-pro1.jpg]]
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As to construct the fusion of DsbA-MBP, a commercial plasmid, pET-39b(+), which contains the gene encoding DsbA, was used as the backbone. The entire coding region of the MBP was amplified by PCR from full length MerR with two pairs of primers. The two PCR products were digested with Xba I / BamH I, or BamH I / Xho I, cloned into Spe I / Xho I digested pET-39b(+) in one step (fig. 1) to make pET-39b(+)-DsbA-MBP.
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[[dsba-pro2.jpg]]
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DsbA and MBD gene was amplified by PCR from pET-39b(+)-DsbA-MBP, with the primer containing T7 polymerase and RBS. The PCR product was digested with EcoR I / Pst I and then cloned into EcoR I / Pst I double digested pSB1K3, to achieve the goal of the standardization of the fusion protein (fig. 2).
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[[dsba-pro3.jpg]]
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As to construct the fusion of DsbA-MerR, a commercial plasmid, pET-39b(+), which contains the gene encoding DsbA, was used as the backbone. The entire coding region of the MerR was amplified by PCR from full length MerR with a pair of primer. The two PCR products were digested with Xba I / Xho I, cloned into Spe I / Xho I digested pET-39b(+) in one step (fig. 3) to make pET-39b(+)-DsbA-MerR.
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[[dsba-pro4.jpg]]
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DsbA and MerR gene was amplified by PCR from pET-39b(+)-DsbA-MerR, with the primer containing T7 polymerase and RBS. The PCR product was digested with EcoR I / Pst I and then cloned into EcoR I / Pst I double digested pSB1K3, to achieve the goal of the standardization of the fusion protein (fig. 4).
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== Results ==
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===User Reviews===
 
===User Reviews===

Revision as of 07:58, 26 October 2010


Applications of BBa_K346033

dsba-pro1.jpg

As to construct the fusion of DsbA-MBP, a commercial plasmid, pET-39b(+), which contains the gene encoding DsbA, was used as the backbone. The entire coding region of the MBP was amplified by PCR from full length MerR with two pairs of primers. The two PCR products were digested with Xba I / BamH I, or BamH I / Xho I, cloned into Spe I / Xho I digested pET-39b(+) in one step (fig. 1) to make pET-39b(+)-DsbA-MBP.

dsba-pro2.jpg


DsbA and MBD gene was amplified by PCR from pET-39b(+)-DsbA-MBP, with the primer containing T7 polymerase and RBS. The PCR product was digested with EcoR I / Pst I and then cloned into EcoR I / Pst I double digested pSB1K3, to achieve the goal of the standardization of the fusion protein (fig. 2).

dsba-pro3.jpg

As to construct the fusion of DsbA-MerR, a commercial plasmid, pET-39b(+), which contains the gene encoding DsbA, was used as the backbone. The entire coding region of the MerR was amplified by PCR from full length MerR with a pair of primer. The two PCR products were digested with Xba I / Xho I, cloned into Spe I / Xho I digested pET-39b(+) in one step (fig. 3) to make pET-39b(+)-DsbA-MerR.

dsba-pro4.jpg

DsbA and MerR gene was amplified by PCR from pET-39b(+)-DsbA-MerR, with the primer containing T7 polymerase and RBS. The PCR product was digested with EcoR I / Pst I and then cloned into EcoR I / Pst I double digested pSB1K3, to achieve the goal of the standardization of the fusion protein (fig. 4).


Results

User Reviews

UNIQee3c4bd1a8f242ff-partinfo-00000000-QINU UNIQee3c4bd1a8f242ff-partinfo-00000001-QINU