Difference between revisions of "Team:Birkbeck/Basic Parts"
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<h2>Basic Parts</h2> | <h2>Basic Parts</h2> | ||
<h3>ORF314 (BBa_K1846000)</h3> | <h3>ORF314 (BBa_K1846000)</h3> | ||
| − | <p>In the commonly used lab strain of bacteriophage | + | <p>In the commonly used lab strain of λ-bacteriophage (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 <i>E.coli</i>. Furthermore, the sequence shows great (>50%) homology with the gene 37 tail fibre protein of bacteriophage T4.</p> |
| − | <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 by restriction with EcoRI and PstI followed by DNA agarose electrophoresis (Fig. 1), with pSB1C3 alone and ORF314 alone as controls. The ORF314 biobrick has been assigned <a href="http://parts.igem.org/Part:BBa_K1846000">BBa_K1846000.</a></p> | + | <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 <i>EcoRI</i> and <i>PstI</i>, before ligating the two pieces together using T4 DNA ligase. Success of the cloning procedure was confirmed by restriction with <i>EcoRI</i> and <i>PstI</i> followed by DNA agarose electrophoresis (<b>Fig. 1</b>), with pSB1C3 alone and ORF314 alone as controls. The ORF314 biobrick has been assigned <a href="http://parts.igem.org/Part:BBa_K1846000">BBa_K1846000.</a></p> |
<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> | <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> | ||
| − | <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 3.</p> | + | <p><b>Fig 1. Agarose gel electrophoresis of ORF314 inserted in pSB1C3 and restricted with <i>EcoRI</i> and <i>PstI</i>.</b> The expected band sizes are 1 kb for ORF314 and 2 kb for pSB1C3. Successful results were obtained for samples 2 and 3.</p> |
Revision as of 21:17, 7 September 2015
Birkbeck iGEM
The Owligos are the first-ever team entered into the international Genetically Engineered Machine (iGEM) Competition by Birkbeck, University of London. We’re a varied group of students who reflect the diversity and unique character of our institution: many of us have chosen science as a second career, having already spent some time in full-time work. For most of us, this has meant making our way through a degree while continuing to work full-time. Hopefully this kind of dedication will help us successfully navigate our way through our iGEM project.
Project Aim
Our project aims to create a new diagnostic solution that will be low-tech and cost-effective enough to allow its usage in deprived and remote communities. We’re attempting to engineer a bacteriophage lambda chassis to change its host affinity, while simultaneously adding a marker that will facilitate easy detection of a target bacterial pathogen in patient samples.
To demonstrate this approach as a proof of concept for the competition, we plan to change this affinity between different strains of E.coli; however, ultimately we hope to demonstrate that this principle could also be applied to alter the phage’s host range to other bacterial species. We could then provide a modular system capable of diagnosing a range of diseases. Of course, we haven’t chosen a simple goal. But as Birkbeck pioneers, we are determined to prove ourselves by making our project a success. We can’t wait to present the results of our work at the Giant Jamboree in September!
Location: Birkbeck, University of London Malet Street London WC1E 7HX Contact us: |
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Tweets by @BBKiGEM |
Our BioBricks
Basic Parts
ORF314 (BBa_K1846000)
In the commonly used lab strain of λ-bacteriophage (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 by restriction with EcoRI and PstI followed by DNA agarose electrophoresis (Fig. 1), with pSB1C3 alone and ORF314 alone as controls. The ORF314 biobrick has been assigned BBa_K1846000.
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 3.