Difference between revisions of "Part:BBa K896000"

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	<partinfo>BBa_K896000 short</partinfo>		<partinfo>BBa_K896000 short</partinfo>
−	SQR introduction:	+
		+
		+	'''SQR introduction''':<p></p>
	Sulfide-dependent anoxygenic photosynthesis, driven by photosystem I (PS/I) alone, among cyanobacteria was first described for Oscillatoria limnetica from Solar Lake. Later photosynthetic sulfide oxidation in O. limnetica led to the discovery of sulfide-quinone reductase (SQR; E.C.1.8.5.′), a novel enzyme that transfers electrons from sulfide into the quinone pool.<p></p>		Sulfide-dependent anoxygenic photosynthesis, driven by photosystem I (PS/I) alone, among cyanobacteria was first described for Oscillatoria limnetica from Solar Lake. Later photosynthetic sulfide oxidation in O. limnetica led to the discovery of sulfide-quinone reductase (SQR; E.C.1.8.5.′), a novel enzyme that transfers electrons from sulfide into the quinone pool.<p></p>
	Above are sulfide-induced sulfide-Quinone Reductase with the electron transport system.<p></p>		Above are sulfide-induced sulfide-Quinone Reductase with the electron transport system.<p></p>
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−	Cloning of SQR gene:	+	'''Gene construction and cloning''':<p></p>
−	SQR gene came from Synechococcus elongatus PCC7002 because the hypersaline strain S. elongatus PCC 7002, which is already sequenced, is 96% similar to O.limnetica SQR gene.<p></p>	+	1. This gene is constructed by http://2012.igem.org/Team:NYMU-Taipei.
−	[[Image:Cloning SQR.jpg]]	+	2. SQR gene came from Synechococcus elongatus PCC7002 because the hypersaline strain S. elongatus PCC 7002, which is already sequenced, is 96% similar to O.limnetica SQR gene. We got the whole gene sequence of SQR from NCBI web  http://www.ncbi.nlm.nih.gov/gene/6054904<p></p>
		+	3. Figure showed the cloning result of SQR gene.<p></p>
		+	Figure 1[[Image:Cloning SQR.jpg]]<p></p>
		+
		+	===Function analysis===
		+
		+	'''Methods and materials'''<p></p>
		+	Details described in https://parts.igem.org/Part:BBa_K896000:Experience.
		+
		+	'''Results'''
		+	After establishing a H2S standard curve to quantify H2S concentration, different H2S concentration challenge SQR transformed E.coli and H2S consumption in 24hrs or 48hrs were tested. The result showed that SQR transformed E.coli consumed much more H2S compred to the blank control, Str transformed E.coli. Our SQR transformed E.coli depleted much more H2S in 48 hours than in 24 hours timepoint.(figure 2A,2B) And SQR transformed E.coli consumed H2S dramatically.(figure 3)<p></p>
		+	Following were results in our experiment.<p></p>
		+
		+	Figure 2<p></p>
		+	(A)different concentration of H2S under 24 hrs[[Image:SQR_24hr.jpg]]<p></p>
		+	(B)different concentration of H2S under 48 hrs[[Image:SQR_48hr.jpg]]<p></p>
		+	Figure 3<p></p>
		+	[[Image:SQR_Na2S500mM.png]]
		+
		+
		+	'''Conclusion'''
		+	Our cloning product SQR(sulfide quinone reductase),from Synechococcus sp. PCC 7002 plasmid pAQ7, could transfer electrons from sulfide into the quinone pool.<p></p>
		+
		+	===Practical Usage===
		+	1. To further practical use, we believe our artificial creatures have high ability to convert hydrogen sulfide to sulfur.
		+	2.Other related parts for solving sulfide problems not only on earth but also on Venus<p></p>
		+
		+	a.[https://parts.igem.org/Part:BBa_K896001 CysI(sulfite reductase)]:BBa_K896001<p></p>
		+
		+	b.[https://parts.igem.org/Part:BBa_K896002 Dsr(sulfite reductase)]:BBa_K896002<p></p>
−	Function analysis of SQR gene:	+	3.We did a great job on sulfur metabolism.Above was the novelty of our engineered E.coli or cyanobacteria.
		+	[[Image:S metabolism.png]]<p></p>
		+	===References===
		+	1.Facultative Anoxygenic Photosynthesis in the Cyanobacterium Oscillatoria limnetica Yehuda Cohen,* Etana Padan, and Moshe Shilo Department of Microbiological Chemistry, The Hebrew University-Hadassah Medical School, Jerusalem, Israel Received for publication 9 May 1975 <p></p>
		+	2.Sulfide oxidation in gram-negative bacteria by expression of the sulfide–quinone reductase gene of Rhodobacter capsulatus and by electron transport to ubiquinone Hiroomi Shibata and Shigeki Kobayashi 2001 <p></p>
		+	3.Sulfur metabolism in Thiorhodoceae. I. Quantitative measurements on growing cells of Chromatium okenii. Antonie Leeuwenhoek, 30: 225–238 Trüper, H.G., and Schlegel, H.G. 1964. <p></p>

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Latest revision as of 16:36, 25 October 2012

SQR(sulfide quinone reductase),from Synechococcus sp. PCC 7002 plasmid pAQ7

SQR introduction:

Sulfide-dependent anoxygenic photosynthesis, driven by photosystem I (PS/I) alone, among cyanobacteria was first described for Oscillatoria limnetica from Solar Lake. Later photosynthetic sulfide oxidation in O. limnetica led to the discovery of sulfide-quinone reductase (SQR; E.C.1.8.5.′), a novel enzyme that transfers electrons from sulfide into the quinone pool.

Above are sulfide-induced sulfide-Quinone Reductase with the electron transport system.

(Cohen, Y., E. Padan, and M. Shilo, Facultative anoxygenic photosynthesis in the cyanobacterium Oscillatoria limnetica. J Bacteriol, 1975. 123(3): p. 855-61.)

Gene construction and cloning:

1. This gene is constructed by http://2012.igem.org/Team:NYMU-Taipei.

2. SQR gene came from Synechococcus elongatus PCC7002 because the hypersaline strain S. elongatus PCC 7002, which is already sequenced, is 96% similar to O.limnetica SQR gene. We got the whole gene sequence of SQR from NCBI web http://www.ncbi.nlm.nih.gov/gene/6054904

3. Figure showed the cloning result of SQR gene.

Figure 1

Function analysis

Methods and materials

Details described in https://parts.igem.org/Part:BBa_K896000:Experience.

Results

After establishing a H2S standard curve to quantify H2S concentration, different H2S concentration challenge SQR transformed E.coli and H2S consumption in 24hrs or 48hrs were tested. The result showed that SQR transformed E.coli consumed much more H2S compred to the blank control, Str transformed E.coli. Our SQR transformed E.coli depleted much more H2S in 48 hours than in 24 hours timepoint.(figure 2A,2B) And SQR transformed E.coli consumed H2S dramatically.(figure 3)

Following were results in our experiment.

Figure 2

(A)different concentration of H2S under 24 hrs

(B)different concentration of H2S under 48 hrs

Figure 3

Conclusion

Our cloning product SQR(sulfide quinone reductase),from Synechococcus sp. PCC 7002 plasmid pAQ7, could transfer electrons from sulfide into the quinone pool.

Practical Usage

1. To further practical use, we believe our artificial creatures have high ability to convert hydrogen sulfide to sulfur.

2.Other related parts for solving sulfide problems not only on earth but also on Venus

a.CysI(sulfite reductase):BBa_K896001

b.Dsr(sulfite reductase):BBa_K896002

3.We did a great job on sulfur metabolism.Above was the novelty of our engineered E.coli or cyanobacteria.

References

1.Facultative Anoxygenic Photosynthesis in the Cyanobacterium Oscillatoria limnetica Yehuda Cohen,* Etana Padan, and Moshe Shilo Department of Microbiological Chemistry, The Hebrew University-Hadassah Medical School, Jerusalem, Israel Received for publication 9 May 1975

2.Sulfide oxidation in gram-negative bacteria by expression of the sulfide–quinone reductase gene of Rhodobacter capsulatus and by electron transport to ubiquinone Hiroomi Shibata and Shigeki Kobayashi 2001

3.Sulfur metabolism in Thiorhodoceae. I. Quantitative measurements on growing cells of Chromatium okenii. Antonie Leeuwenhoek, 30: 225–238 Trüper, H.G., and Schlegel, H.G. 1964.

Sequence and Features