Difference between revisions of "Team:NYMU-Taipei/Pigments"

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<p>  Melanin, a biological pigment widely found in terrestrial flora and fauna, is a complex oxidation product of amino acid tyrosine. Melanin plays diverse roles in a myriad of organisms. As far as the human is concerned, melanin is the primary determinant of human skin color and pupils or irises of the eyes. It is also an important signal molecule in the human neural system. For microorganisms, melanin would protect them by against ultraviolet radiation effect from sunlight, which is detrimental to most of the organisms.</p>
 
<p>  Melanin, a biological pigment widely found in terrestrial flora and fauna, is a complex oxidation product of amino acid tyrosine. Melanin plays diverse roles in a myriad of organisms. As far as the human is concerned, melanin is the primary determinant of human skin color and pupils or irises of the eyes. It is also an important signal molecule in the human neural system. For microorganisms, melanin would protect them by against ultraviolet radiation effect from sunlight, which is detrimental to most of the organisms.</p>
<p>  Due to the fact that the dark pigment derived from <font class='mark_black'>MelA gene has extensive wavelength absorbance</font>, we decided to transform MelA gene combined with particular constitute promoter into <i>Synechococcus elongates</i> PCC 7942 to measure the growth curve and photosynthetic efficiency of it. Based on the well-elaborated procedure provide by IGEM Tokyo Tech 2009<sup>9</sup>, we had intended to replicate their experiment to produce melanin massively first in <i>E. coli</i>, but failed to succeed due to inconsistence of DNA sequence. Therefore, we had no alternative but to turn to DNA synthesis and directly transform MelA gene ligated with our backbone into microalgae.</p>
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<p>  Due to the fact that the dark pigment derived from <font class='mark_black'>MelA gene has extensive wavelength absorbance</font>, we decided to transform MelA gene combined with particular constitute promoter into <i>Synechococcus elongatus</i> PCC 7942 to measure the growth curve and photosynthetic efficiency of it. Based on the well-elaborated procedure provide by IGEM Tokyo Tech 2009<sup>9</sup>, we had intended to replicate their experiment to produce melanin massively first in <i>E. coli</i>, but failed to succeed due to inconsistence of DNA sequence. Therefore, we had no alternative but to turn to DNA synthesis and directly transform MelA gene ligated with our backbone into microalgae.</p>
 
<center><img src='https://static.igem.org/mediawiki/2017/8/8e/T--NYMU-Taipei--pigments_melanin.png' style='width:70%'></center>
 
<center><img src='https://static.igem.org/mediawiki/2017/8/8e/T--NYMU-Taipei--pigments_melanin.png' style='width:70%'></center>
 
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<p>  pPIGBACK-CrtZ is transformed into <i>Synechoccocus elongatus</i> PCC7942 after 20 days cultivation, the transformants electrophoresis result is showed below. <font class='mark_backbone'>Transformation efficiency</font> of pPIGBACK-CrtZ is 11.4 transformants per μg DNA, and correctness is 52% (10/19), which is relatively high compared to low successful rate of gene recombination.</p>
 
<p>  pPIGBACK-CrtZ is transformed into <i>Synechoccocus elongatus</i> PCC7942 after 20 days cultivation, the transformants electrophoresis result is showed below. <font class='mark_backbone'>Transformation efficiency</font> of pPIGBACK-CrtZ is 11.4 transformants per μg DNA, and correctness is 52% (10/19), which is relatively high compared to low successful rate of gene recombination.</p>
<p>  Therefore, we can conclude that pPIGBACK is quite a reliable vector which could finish gene double-crossover homologous recombination in <i>S. elongates</i> PCC 7942 genome, because the successful rate of gene double-crossingover homologous recombination is low in cyanobacteria. Moreover, compare pPIGBACK-CrtZ transformants with wild type, we can assure that pPIGBACK could be express in <i>S. elongates</i> PCC 7942, which is such a milestone in our project. With <font class='mark_backbone'>the high correctness of pPIGBACK-CrtZ transformants</font>, we have the confidence that multiple colors of cyanobacteria is possible and could be functional in the near future.</p>
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<p>  Therefore, we can conclude that pPIGBACK is quite a reliable vector which could finish gene double-crossover homologous recombination in <i>S. elongatus</i> PCC 7942 genome, because the successful rate of gene double-crossingover homologous recombination is low in cyanobacteria. Moreover, compare pPIGBACK-CrtZ transformants with wild type, we can assure that pPIGBACK could be express in <i>S. elongatus</i> PCC 7942, which is such a milestone in our project. With <font class='mark_backbone'>the high correctness of pPIGBACK-CrtZ transformants</font>, we have the confidence that multiple colors of cyanobacteria is possible and could be functional in the near future.</p>
 
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<img src="https://static.igem.org/mediawiki/2017/8/80/T--NYMU-Taipei--pigment_CrtZ.jpg" style="width:95%">
 
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<li>Copeland,A. et al.(2014). Carotenoid pathway of <i>Synechococcus elongates</i> PCC 7942, complete genomegatus PCC 7942. Unpublished.
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<li>Copeland,A. et al.(2014). Carotenoid pathway of <i>Synechococcus elongatus</i> PCC 7942, complete genomegatus PCC 7942. Unpublished.
 
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Revision as of 13:34, 30 October 2017

Pigments

  In our project, we transfer five types of pigment-related gene sequence (Indigoidine, Zeaxanthin, Melanin, Astaxanthin and Lycopene) into our cyanobacteria. We expect to get five different colors of microalgae, so we could see whether changing the original color of microalgae would change wavelength absorbance and have better photosynthetic efficiencies. Due to better photosynthetic efficiencies, we could elevate oil accumulation in microalgae, which would have great benefit in both industry and scientific usage.