Difference between revisions of "Team:Birkbeck/Basic Part"

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<h1>Under Construction</h1>
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<h1>Our BioBricks</h1>
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<h2> Basic Parts</h2>
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<h2>Basic Parts</h2>
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<h3>ORF314 (BBa_K1846000)</h3>
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<p>In the commonly used lab strain of bacteriophage Lambda (known as λPaPa), ORF314 is one of two open reading frames resulting from a frameshift in the tail fibre protein gene (stf). This ORF codes for the C-terminus of the protein, which contains the host receptor recognition site. ORF314 binds to the OmpC protein of E.coli. Furthermore, the sequence shows great (>50%) homology with the gene 37 tail fibre protein of bacteriophage T4.</p>
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<p>To transform this sequence into a BioBrick basic part, we had the coding sequence synthesised, including the BioBrick prefix and suffix and an additional section to facilitate the cloning of complementary sequence ORF401. We then restricted both our synthesised gene and the linearised plasmid backbones (pSB1C3 for shipping and pSB1K3 for further processing) with EcoRI and PstI, before ligating the two pieces together using T4 DNA ligase. Success of the cloning procedure was confirmed restriction with EcoRI and PstI followed by DNA agarose electrophoresis (Fig. 1), with pSB1C3 alone and ORF314 alone as controls.</p>
  
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<a href='http://postimage.org/' target='_blank'><img src='http://s23.postimg.org/8de2ulxyz/150730_ORF314_p_SB1_C3_annotated.jpg' border='0' alt="Agarose gel electrophoresis of ORF314 in vector" /></a><br /><a target='_blank' href='http://postimage.org/'></a><br /><br />
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<p><b>Fig 1. Agarose gel electrophoresis of ORF314 inserted in pSB1C3 and restricted with EcoRI and PstI.</b> The expected band sizes are 1 kb for ORF314 and 2 kb for pSB1C3. Successful results were obtained for samples 2 and 4.</p>
  
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<p>In order to be considered for the <a href="https://2015.igem.org/Judging/Awards#SpecialPrizes">Best New Basic Part award</a>, you must fill out this page. Please give links to the Registry entries for the Basic parts you have made. Please see the Registry's <a href="http://parts.igem.org/Help:Parts#Basic_and_Composite_Parts"> Help:Parts page</a> for more information on part types.</p>
 
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A <b>basic part</b> is a functional unit of DNA that cannot be subdivided into smaller component parts. <a href="http://parts.igem.org/wiki/index.php/Part:BBa_R0051">BBa_R0051</a> is an example of a basic part, a promoter regulated by lambda cl.
 
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<p>Most genetically-encoded functions have not yet been converted to BioBrick parts. Thus, there are <b>many</b> opportunities to find new, cool, and important genetically encoded functions, and refine and convert the DNA encoding these functions into BioBrick standard biological parts. </p>
 
  
 
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Revision as of 13:19, 31 July 2015

Our BioBricks

Basic Parts

ORF314 (BBa_K1846000)

In the commonly used lab strain of bacteriophage Lambda (known as λPaPa), ORF314 is one of two open reading frames resulting from a frameshift in the tail fibre protein gene (stf). This ORF codes for the C-terminus of the protein, which contains the host receptor recognition site. ORF314 binds to the OmpC protein of E.coli. Furthermore, the sequence shows great (>50%) homology with the gene 37 tail fibre protein of bacteriophage T4.

To transform this sequence into a BioBrick basic part, we had the coding sequence synthesised, including the BioBrick prefix and suffix and an additional section to facilitate the cloning of complementary sequence ORF401. We then restricted both our synthesised gene and the linearised plasmid backbones (pSB1C3 for shipping and pSB1K3 for further processing) with EcoRI and PstI, before ligating the two pieces together using T4 DNA ligase. Success of the cloning procedure was confirmed restriction with EcoRI and PstI followed by DNA agarose electrophoresis (Fig. 1), with pSB1C3 alone and ORF314 alone as controls.

Agarose gel electrophoresis of ORF314 in vector


Fig 1. Agarose gel electrophoresis of ORF314 inserted in pSB1C3 and restricted with EcoRI and PstI. The expected band sizes are 1 kb for ORF314 and 2 kb for pSB1C3. Successful results were obtained for samples 2 and 4.