Difference between revisions of "Team:NWU-CHINA/Demonstrate"

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  <h1>Preparation</h1>
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<h3>Acquire sample</h3>
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<p>We got crude oil from Yanchang oil field production of Xiasiwan (36°27′36″ N, 108°55′12″ E), a terrestrial oil production located in Northern Shaanxi, China for decades. Petroleum hydrocarbons con- taminated soils were also collected from Yanchang oil field production of Xiasiwan. The soils were taken from 2 to10 cm depth layer and sieved through a 2-mm screen, then sample was stored at 5 °C in the dark until required. </p>
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<h3>Sample separation </h3>
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<p>We separate a strain capable of efficiently degrading alkanes from samples. </p>
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<p>There are many bacterial strains isolated from petroleum-contaminated soil which had the ability to either produce visual rhamnolipids as indicated by the presence of blue halos, or utilize crude oil as a sole source of carbon and energy. Among these isolates, one strain code as DN1 was not only capable of highly producing the biosurfactants, but it was also able to degrade crude oil effectively. Then the complete 16SrRNA gene sequence (1475bp) of strain DN1 was obtained and phylogenetic analysis was conducted to determine that it belonged to Gamma Proteobacteria, showing the highest 16SrRNA gene sequence similarity of 98% with Pseudomonas aeruginosa. This strain was oval to rod-shaped (0.5–0.8 mm_1.5–3.0 mm), gram-negative and motile with a single polar flagellum (Figure 1). Colonies(2mm–3 mm)growing on LB agar for 24h at 37℃ were rough, circular convex, wet and yellowish-brown in color (Figure 3). The strain was tolerant to a wide variety of  physical conditions, including temperature and pH, with the optimum growth occurring at a pH6.5–7.5 and 30–37°C. It was positive for citrate, catalase, oxidase, aerobic nitrite reduction, anaerobic nitrate reduction and denitrification, as well as for the hydrolysis of gelatin. According to API-test results, the biochemical profiles seemed to ferment some carbohydrates by the strain. Morphological, physiological and phylogenetic properties indicated that strainDN1(CCTCCNO:M2011287)was a member of the genus Pseudomonas, and the accession number in the GenBank is KP119458. </p>
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        <div align="center"><img src="https://static.igem.org/mediawiki/2017/f/f6/NWU-CHINA_Figure1.png"/ width="70%"></div>
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    <div style="text-align:center">Figure1. Phylogenetic relationship between strain DN1 and species in the Pseudomonas based on the 16S rRNA gene sequences, constructed by Neighbor-Joining algorithm.</div>
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    <div style="text-align:center">Figure2. DN1 strain under optical microscope after Gram stainning.</div>
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<div style="text-align:center">Figure3. DN1 strain colonies on plate.</div>
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Reference
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<br>[1] Kuang-Yi Ma, Meng-Yan Sun, Wen Dong, Chun-Qiu He, Fu-Lin Chen, Yan-Ling Ma .Effects of nutrition optimization strategy on rhamnolipid production in a Pseudomonas aeruginosa strain DN1 for bioremediation of crude oil[J].Biocatalysis and Agricultural Biotechnology, 2016, 6(1):144-151.
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<br>[2] Wen Dong, Chunqiu He, Yanpeng Li, Chao Huang, Fulin Chen, Yanling Ma⁎.Complete genome sequence of a versatile hydrocarbon degrader, Pseudomonas aeruginosa DN1 isolated from petroleum-contaminated soil[J].Gene Reports, 2017, 7(1):123-126.
  
  
 
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<h3>★  ALERT! </h3>
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<p>This page is used by the judges to evaluate your team for the <a href="https://2017.igem.org/Judging/Medals">medal criterion</a> or <a href="https://2017.igem.org/Judging/Awards"> award listed above</a>. </p>
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<p> Delete this box in order to be evaluated for this medal criterion and/or award. See more information at <a href="https://2017.igem.org/Judging/Pages_for_Awards"> Instructions for Pages for awards</a>.</p>
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&copy;2017 NWU-CHINA IGEM.All Rights Reserved. <br>
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<h1>Demonstrate</h1>
 
<h3>Gold Medal Criterion #4</h3>
 
 
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Teams that can show their system working under real world conditions are usually good at impressing the judges in iGEM. To achieve gold medal criterion #4, convince the judges that your project works. There are many ways in which your project working could be demonstrated, so there is more than one way to meet this requirement. This gold medal criterion was introduced in 2016, so check our what 2016 teams did to achieve a their gold medals!
 
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Please see the <a href="https://2017.igem.org/Judging/Medals">2017 Medals Page</a> for more information.
 
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<h4> What should we do for our demonstration?</h4>
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<h5> Standard teams </h5>
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If you have built a proof of concept system, you can demonstrate it working under real world conditions. If you have built a biological device that is intended to be a sensor, can you show it detecting whatever it is intended to sense. If it is intended to work in the field, you can show how this might work using a simulated version in the lab, or a simulation of your device in the field.<strong> Please note biological materials must not be taken out of the lab</strong>.
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<h5> Special track teams </h5>
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Special track teams can achieve this medal criterion by bringing their work to the Jamboree and showcasing it in the track event. Art & Design, Measurement, Hardware and Software tracks will all have showcase events at the Giant Jamboree.<strong> Please note biological materials must not be taken out of the lab</strong>.
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Revision as of 07:25, 27 October 2017

Preparation

Acquire sample

We got crude oil from Yanchang oil field production of Xiasiwan (36°27′36″ N, 108°55′12″ E), a terrestrial oil production located in Northern Shaanxi, China for decades. Petroleum hydrocarbons con- taminated soils were also collected from Yanchang oil field production of Xiasiwan. The soils were taken from 2 to10 cm depth layer and sieved through a 2-mm screen, then sample was stored at 5 °C in the dark until required.

Sample separation

We separate a strain capable of efficiently degrading alkanes from samples.

There are many bacterial strains isolated from petroleum-contaminated soil which had the ability to either produce visual rhamnolipids as indicated by the presence of blue halos, or utilize crude oil as a sole source of carbon and energy. Among these isolates, one strain code as DN1 was not only capable of highly producing the biosurfactants, but it was also able to degrade crude oil effectively. Then the complete 16SrRNA gene sequence (1475bp) of strain DN1 was obtained and phylogenetic analysis was conducted to determine that it belonged to Gamma Proteobacteria, showing the highest 16SrRNA gene sequence similarity of 98% with Pseudomonas aeruginosa. This strain was oval to rod-shaped (0.5–0.8 mm_1.5–3.0 mm), gram-negative and motile with a single polar flagellum (Figure 1). Colonies(2mm–3 mm)growing on LB agar for 24h at 37℃ were rough, circular convex, wet and yellowish-brown in color (Figure 3). The strain was tolerant to a wide variety of physical conditions, including temperature and pH, with the optimum growth occurring at a pH6.5–7.5 and 30–37°C. It was positive for citrate, catalase, oxidase, aerobic nitrite reduction, anaerobic nitrate reduction and denitrification, as well as for the hydrolysis of gelatin. According to API-test results, the biochemical profiles seemed to ferment some carbohydrates by the strain. Morphological, physiological and phylogenetic properties indicated that strainDN1(CCTCCNO:M2011287)was a member of the genus Pseudomonas, and the accession number in the GenBank is KP119458.

Figure1. Phylogenetic relationship between strain DN1 and species in the Pseudomonas based on the 16S rRNA gene sequences, constructed by Neighbor-Joining algorithm.
Figure2. DN1 strain under optical microscope after Gram stainning.
Figure3. DN1 strain colonies on plate.



Reference
[1] Kuang-Yi Ma, Meng-Yan Sun, Wen Dong, Chun-Qiu He, Fu-Lin Chen, Yan-Ling Ma .Effects of nutrition optimization strategy on rhamnolipid production in a Pseudomonas aeruginosa strain DN1 for bioremediation of crude oil[J].Biocatalysis and Agricultural Biotechnology, 2016, 6(1):144-151.
[2] Wen Dong, Chunqiu He, Yanpeng Li, Chao Huang, Fulin Chen, Yanling Ma⁎.Complete genome sequence of a versatile hydrocarbon degrader, Pseudomonas aeruginosa DN1 isolated from petroleum-contaminated soil[J].Gene Reports, 2017, 7(1):123-126.

©2017 NWU-CHINA IGEM.All Rights Reserved.