Difference between revisions of "Part:BBa K1998003"
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<partinfo>BBa_K1998003 parameters</partinfo>
 
<partinfo>BBa_K1998003 parameters</partinfo>
 
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===Overview===
 
===Overview===
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This biobrick contains one gene from the photosystem II complex of Chlamydomonas reinhardtii - psbD. It is driven by a Plac promoter at the beginning of the sequence, with a RBS preceding the gene, and terminator sequence at the 3' end. The psbD gene that comprises this part encodes the D2 protein, which is one of the two proteins that comprise the Photosystem II reaction center core.
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This biobrick contains one gene from the photosystem II complex of Chlamydomonas reinhardtii - <i>psbD</i>. It is driven by a Plac promoter at the beginning of the sequence, with a RBS preceding the gene, and terminator sequence at the 3' end. The <i>psbD</i> gene that comprises this part encodes the D2 protein, which is one of the two proteins that comprise the Photosystem II reaction center core.
 
<br><br>
 
<br><br>
 
<html><center><img src= "https://static.igem.org/mediawiki/2016/a/aa/T--Macquarie_Australia--PhotosystemIISynthesis.png" alt="PhotosystemIISynthesis" height="50%"width="75%"></center></html>
 
<html><center><img src= "https://static.igem.org/mediawiki/2016/a/aa/T--Macquarie_Australia--PhotosystemIISynthesis.png" alt="PhotosystemIISynthesis" height="50%"width="75%"></center></html>
  
 
===Biology & Literature===
 
===Biology & Literature===
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Hydrogenase 1 (Hyd1) is a gene which codes for a peptide containing 10 D-amino acids which has been known to induce cell autophagy resulting from an increase in reactive oxygen species and ATP depletion of the cell [1]. Hyd1 has also been known to help treat prostate cancer by preventing the adhesion of carcinomas to the epithelium lining of the digestive tract [2].
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This biobrick contains one gene from the photosystem II complex of <i>Chlamydomonas reinhardtii</i> - <i>psbD</i>. It is driven by a Plac promoter at the beginning of the sequence, with a RBS preceding the gene, and terminator sequence at the 3' end. The <i>psbD</i> gene that comprises this part encodes the D2 protein, which is one of the two proteins that comprise the Photosystem II reaction center core P680 [1, 2]. It has been shown that the <i>psbD</i> gene is highly conserved within <i>C. rienhardtii</i> [3].
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<br><br>
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<br>
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Mutations in the <i>psbD</i> gene causes the D2 peptide to not be visible when screened perhaps indicating it's instability within the complex [3]. In addition it was found that other proteins are affected by a mutation in the <i>psbD</i> gene due to the lack of accumulation of other PSII proteins. Therefore the role of <i>psbD</i> is to provide stability in the membrane of the complex as well as to regulate the expression of the D1 protein of the complex [3].
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More specifically Hyd1 catalyses the oxidation of hydrogen in the anaerobic respiration of <i>E.coli</i> this done by working in parallel with Hyd 2 before fermentation is undertaken by Hyd3. Hyd1 is a membrane bound hydrogenase that facilitates the uptake of oxidated hydrogen gas [3]. The hya operon encodes the Hydrogenase peptide which is initiated under anaerobic conditions as well as acidic pH levels which signals that H+ must be facilitated to move from ICF to ECF and viceverser to maintain pH levels in <i>E.coli</i> [4].
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===Part Verification===
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<html><centre><img src=" https://static.igem.org/mediawiki/2016/8/82/T--Macquarie_Australia--PSII_Show_Gel_and_.JPG " height="20%" width="40%"></center></html>
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<br><br>
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<b>Fig 1.</b> Gel electrophoresis of the operons and single parts constituting the Photosystem II pathway implemented in this project. The part psbD (1210 bp) can be seen in lane 6 on the gel at the correct size relative to the 1kb ladder and other parts in our project shown on this gel. The band higher up in this lane on the gel is the pSB1C3 backbone (2000 bp).
  
 
===Protein information===
 
===Protein information===
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Protein sequence:  
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<i>psbD</i><br>
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MSALVLKPCAAVSIRGSSCRARQVAPRAPLAASTVRVALATLEAPARRLGNVACAAAAPAAEAPLSHVQQALAELAKPKDDPTRKHVCVQVAPAVRVAIAETLGLAPGATT
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Mass: 43kDa<br>
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PKQLAEGLRRLGFDEVFDTLFGADLTIMEEGSELLHRLTEHLEAHPHSDEPLPMFTSCCPGWIAMLEKSYPDLIPYVSSCKSPQMMLAAMVKSYLAEKKGIAPKDMVMV
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Sequence: <br>
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SIMPCTRKQSEADRDWFCVDADPTLRQLDHVITTVELGNIFKERGINLAELPEGEWDNPMGVGSGAGVLFGTTGGVMEAALRTAYELFTGTPLPRLSLSEVRGMDGIKET
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LEGFTLYASGSYVVWKEVKKQLRSVHIKRNAHGSMTLMTGFVKTVSYSVGQVYYYSLVLTLHVVGLVLLSLLHGIRMVLLLTKVVTSQQLFLHLLTVWLTLFYLFGVQKLKVI
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NITMVPAPGSKFEELLKHRAAARAEAAAHGTPGPLAWDGGAGFTSEDGRGGITLRVAVANGLGNAKKLITKMQAGEAKYDFVEIMACPAGCVGGGGQPRSTDKAITQKR
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SLVGVNLVVYGHSLLYTVHLVLVSCFVSLKLLVQTYVHTTQLLSQHQLLYSFQYSFTHVNQVGSLHLVSVLLSSVSFYSSKVSTTGHLTHSTWVLLVFVLLYYVLFTVLLLKTH
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QAALYNLDEKSTLRRSHENPSIRELYDTYLGEPLGHKAHELLHTHYVAGGVEEKDEKK
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YSKTVTVLTHSVHSTLHRLKKHTLWLLLTVSGHKSSVLLSLTNVGFTSSCYFQLVFGVLLVLVLTYVLTTSYHKRFVLLKNPEFETFYTKKHSSRKVFLAWKGCFRTNHTKRL
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VFSKKIYHRGKPSYITNKRRLAIPGKKRARGSHPRALFFQKX
 
<br>
 
<br>
  
 
===References===
 
===References===
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[1]
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[1] Kim M, Christopher DA, Mullet JE. ADP-Dependent Phosphorylation Regulates Association of a DNA-Binding Complex with the Barley Chloroplast <i>psbD</i> Blue-Light-Responsive Promoter. Plant physiology. 1999 Feb 1;119(2):663-70.
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<br>
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<br><br>
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[2]
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[2] Rasmussen OF, Bookjans G, Stummann BM, Henningsen KW. Localization and nucleotide sequence of the gene for the membrane polypeptide D2 from pea chloroplast DNA. Plant molecular biology. 1984 Jul 1;3(4):191-9.
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<br>
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<br><br>
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[3]
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[3] Erickson JM, Rahire M, Malnoë P, Girard-Bascou J, Pierre Y, Bennoun P, Rochaix JD. Lack of the D2 protein in a Chlamydomonas reinhardtii <i>psbD</i> mutant affects photosystem II stability and D1 expression. The EMBO journal. 1986 Aug;5(8):1745.
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<br>
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[4]
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<br>
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Latest revision as of 03:42, 21 October 2016


psbD

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Overview

This biobrick contains one gene from the photosystem II complex of Chlamydomonas reinhardtii - psbD. It is driven by a Plac promoter at the beginning of the sequence, with a RBS preceding the gene, and terminator sequence at the 3' end. The psbD gene that comprises this part encodes the D2 protein, which is one of the two proteins that comprise the Photosystem II reaction center core.

PhotosystemIISynthesis

Biology & Literature

This biobrick contains one gene from the photosystem II complex of Chlamydomonas reinhardtii - psbD. It is driven by a Plac promoter at the beginning of the sequence, with a RBS preceding the gene, and terminator sequence at the 3' end. The psbD gene that comprises this part encodes the D2 protein, which is one of the two proteins that comprise the Photosystem II reaction center core P680 [1, 2]. It has been shown that the psbD gene is highly conserved within C. rienhardtii [3].

Mutations in the psbD gene causes the D2 peptide to not be visible when screened perhaps indicating it's instability within the complex [3]. In addition it was found that other proteins are affected by a mutation in the psbD gene due to the lack of accumulation of other PSII proteins. Therefore the role of psbD is to provide stability in the membrane of the complex as well as to regulate the expression of the D1 protein of the complex [3].

Part Verification



Fig 1. Gel electrophoresis of the operons and single parts constituting the Photosystem II pathway implemented in this project. The part psbD (1210 bp) can be seen in lane 6 on the gel at the correct size relative to the 1kb ladder and other parts in our project shown on this gel. The band higher up in this lane on the gel is the pSB1C3 backbone (2000 bp).

Protein information

psbD
Mass: 43kDa
Sequence:
LEGFTLYASGSYVVWKEVKKQLRSVHIKRNAHGSMTLMTGFVKTVSYSVGQVYYYSLVLTLHVVGLVLLSLLHGIRMVLLLTKVVTSQQLFLHLLTVWLTLFYLFGVQKLKVI SLVGVNLVVYGHSLLYTVHLVLVSCFVSLKLLVQTYVHTTQLLSQHQLLYSFQYSFTHVNQVGSLHLVSVLLSSVSFYSSKVSTTGHLTHSTWVLLVFVLLYYVLFTVLLLKTH YSKTVTVLTHSVHSTLHRLKKHTLWLLLTVSGHKSSVLLSLTNVGFTSSCYFQLVFGVLLVLVLTYVLTTSYHKRFVLLKNPEFETFYTKKHSSRKVFLAWKGCFRTNHTKRL VFSKKIYHRGKPSYITNKRRLAIPGKKRARGSHPRALFFQKX

References

[1] Kim M, Christopher DA, Mullet JE. ADP-Dependent Phosphorylation Regulates Association of a DNA-Binding Complex with the Barley Chloroplast psbD Blue-Light-Responsive Promoter. Plant physiology. 1999 Feb 1;119(2):663-70.

[2] Rasmussen OF, Bookjans G, Stummann BM, Henningsen KW. Localization and nucleotide sequence of the gene for the membrane polypeptide D2 from pea chloroplast DNA. Plant molecular biology. 1984 Jul 1;3(4):191-9.

[3] Erickson JM, Rahire M, Malnoë P, Girard-Bascou J, Pierre Y, Bennoun P, Rochaix JD. Lack of the D2 protein in a Chlamydomonas reinhardtii psbD mutant affects photosystem II stability and D1 expression. The EMBO journal. 1986 Aug;5(8):1745.