Difference between revisions of "Team:Worldshaper-Nanjing/Composite Part"

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                                 <h2>COMMON Page</h2>
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                                 <h2>INP-FBPA</h2>
 
                             </div>
 
                             </div>
 
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                             <div class="nj-box4">
 
                                 <p>
 
                                 <p>
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                                     Lorem ipsum dolor sit amet, consectetur adipisicing elit. Quas sit expedita, iusto repellendus cumque, officia architecto consequatur illo fuga eum sed ut autem eos voluptas. Nemo, a, rem! Atque quisquam aperiam eaque tenetur autem, soluta itaque omnis. Minus nesciunt, sint, animi illum quo ab voluptate esse delectus unde maiores iure, quasi a suscipit ipsam aliquid voluptatem. Perspiciatis eveniet, pariatur illum aut cum dolor neque consequatur error aliquid facilis in quasi temporibus assumenda tempore, doloremque autem saepe enim nihil. Voluptates asperiores ullam voluptate quas similique ratione quia hic, eum distinctio laboriosam, consectetur tempora voluptatibus optio natus cumque est necessitatibus dolore alias.
+
                                     BBa_K2513034
 
                                 </p>
 
                                 </p>
 
                                 <p>
 
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                                     Lorem ipsum dolor sit amet, consectetur adipisicing elit. Veritatis expedita repellendus laboriosam aliquid. Neque doloribus ea, id reprehenderit alias saepe debitis eligendi molestias odit, nesciunt rem. Dolorem saepe, provident dolore nesciunt laudantium nostrum enim natus veritatis harum maxime et iure ratione, nulla. Minus excepturi commodi tempore voluptate. Blanditiis similique dolor asperiores ex excepturi perspiciatis, dolores id esse. Voluptate beatae nesciunt cum esse ratione officiis necessitatibus blanditiis ea, laboriosam fugit vero maxime? Voluptatum illo dolorum autem pariatur quisquam. Voluptates soluta culpa necessitatibus veritatis tempora incidunt doloribus placeat repellat et facilis eum sapiente fugit numquam aut, laboriosam aspernatur, esse, magnam excepturi repudiandae amet voluptas nulla quidem. Veritatis nisi consequuntur saepe qui quisquam dignissimos assumenda, iusto odio. Dignissimos reprehenderit esse iusto cupiditate nisi enim, animi similique itaque, perspiciatis error qui. Aperiam, architecto provident.
+
                                     INP-FBPA
 
                                 </p>
 
                                 </p>
 +
                                <p>This part encodes a fusion protein that composed of INP and FBPA and designed to be expressed at the surface of E.coli or Synechococcus sp. PCC 7002. We replaced the 6His tag of INP-6His (BBa_K2509019) with a ferric iron binding protein FBPA (BBa_K2513032)to build this part.</p>
 +
                                <p>BBa_K2509019 and BBa_K2513032</p>
 +
                                <p>We directly replaced the 6His tag with FBPA without being deleted its stop codon to make a C-fusion protein. So that the whole part can only be transferred as a fusion protein with N part of INP and FBPA.</p>
 +
                                <p>Results:</p>
 +
                                <p>Constructions and expression results</p>
 +
                                <p>Metal binding result 1</p>
 +
                                <p>Metal binding result 2</p>
 +
 +
                                <p>cpcBA promoter</p>
 +
                                <p>BBa_K2513032</p>
 +
                                <p>cpcBA promoter</p>
 +
                                <p>This basic part was designed to be ligated into a basic vector pAQ1Basic3 to make an expression vector pAQ1J2 and functioned as a promoter to driver transcription of the target genes downstream. The cpcBA promoter is originally from Synechocystis sp. PCC 6803 and also works well in Synechococcus sp. PCC 7002.</p>
 +
                                <p>The sequence is from Synechocystis sp. PCC 6803, and synthesized by GenScript Corporation.</p>
 +
                                <p>This promoter region can be easily replace by others by digestion with KpnI and NdeI restriction enzymes. So we can easily compare and measure different promoters in Synechococcus sp. PCC 7002 when inserting a reporter gene like EGFP.</p>
 +
 +
                                <p>GlgA2</p>
 +
                                <p>BBa_K2513020</p>
 +
                                <p>GlgA2</p>
 +
                                <p>This part encodes a protein GlgA2 of Synechocystis sp. PCC 6803 that involved in the pathway glycogen biosynthesis, which is part of glycan biosynthesis. We want to increase the starch accumulation of Synechococcus sp. PCC 7002 by transferring this gene.</p>
 +
                                <p>The amino acid sequence is according to the GlgA2 gene of Synechocystis sp. PCC 6803, and the DNA fragment is synthesized by GenScript Corporation.</p>
 +
 +
                                <p>SmtA-out</p>
 +
                                <p>BBa_K2513031</p>
 +
                                <p>SmtA-out</p>
 +
 +
                                <p>This encodes a protein SmtA from Synechococcus sp.PCC7942 which may play a role in essential metal ion homeostasis (especially zinc homeostasis) and resistance to certain non-essential metal ions. Metallothioneins have a high content of cysteine residues that bind various heavy metals. It was designed to be fused with INP or SomA which are well verified proteins that used in surface-display in E.coli and Synechococcus sp. PCC 7002, so that it can be also expressed at the surface of E.coli or Synechococcus sp. PCC 7002 to help the transgenetic hosts to absorbing metal irons.</p>
 +
 +
                                <p>This part can be easily fused with other outer membrane proteins like INP or SomA with or without stop codon using unique restriction sites of BamHI and HindIII at the two sides.</p>
 +
 +
                                <p>The amino sequence is according to the SmtA gene of Synechococcus sp.PCC7942, and DNA fragment is synthesized by GenScript Corporation.</p>
 +
 
                             </div>
 
                             </div>
 
                         </div>
 
                         </div>

Revision as of 11:24, 30 October 2017

INP-FBPA

BBa_K2513034

INP-FBPA

This part encodes a fusion protein that composed of INP and FBPA and designed to be expressed at the surface of E.coli or Synechococcus sp. PCC 7002. We replaced the 6His tag of INP-6His (BBa_K2509019) with a ferric iron binding protein FBPA (BBa_K2513032)to build this part.

BBa_K2509019 and BBa_K2513032

We directly replaced the 6His tag with FBPA without being deleted its stop codon to make a C-fusion protein. So that the whole part can only be transferred as a fusion protein with N part of INP and FBPA.

Results:

Constructions and expression results

Metal binding result 1

Metal binding result 2

cpcBA promoter

BBa_K2513032

cpcBA promoter

This basic part was designed to be ligated into a basic vector pAQ1Basic3 to make an expression vector pAQ1J2 and functioned as a promoter to driver transcription of the target genes downstream. The cpcBA promoter is originally from Synechocystis sp. PCC 6803 and also works well in Synechococcus sp. PCC 7002.

The sequence is from Synechocystis sp. PCC 6803, and synthesized by GenScript Corporation.

This promoter region can be easily replace by others by digestion with KpnI and NdeI restriction enzymes. So we can easily compare and measure different promoters in Synechococcus sp. PCC 7002 when inserting a reporter gene like EGFP.

GlgA2

BBa_K2513020

GlgA2

This part encodes a protein GlgA2 of Synechocystis sp. PCC 6803 that involved in the pathway glycogen biosynthesis, which is part of glycan biosynthesis. We want to increase the starch accumulation of Synechococcus sp. PCC 7002 by transferring this gene.

The amino acid sequence is according to the GlgA2 gene of Synechocystis sp. PCC 6803, and the DNA fragment is synthesized by GenScript Corporation.

SmtA-out

BBa_K2513031

SmtA-out

This encodes a protein SmtA from Synechococcus sp.PCC7942 which may play a role in essential metal ion homeostasis (especially zinc homeostasis) and resistance to certain non-essential metal ions. Metallothioneins have a high content of cysteine residues that bind various heavy metals. It was designed to be fused with INP or SomA which are well verified proteins that used in surface-display in E.coli and Synechococcus sp. PCC 7002, so that it can be also expressed at the surface of E.coli or Synechococcus sp. PCC 7002 to help the transgenetic hosts to absorbing metal irons.

This part can be easily fused with other outer membrane proteins like INP or SomA with or without stop codon using unique restriction sites of BamHI and HindIII at the two sides.

The amino sequence is according to the SmtA gene of Synechococcus sp.PCC7942, and DNA fragment is synthesized by GenScript Corporation.

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