Team:Yale/Our Project/Methods
From 2010.igem.org
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Our plasmid is composed of three parts: a promoter and a terminator Biobrick as well as a novel addition to the biobrick library, the phsABC gene that is known to encode Thiosulfate Reductase. | Our plasmid is composed of three parts: a promoter and a terminator Biobrick as well as a novel addition to the biobrick library, the phsABC gene that is known to encode Thiosulfate Reductase. | ||
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(1) phsABC gene and vector | (1) phsABC gene and vector | ||
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<div id="caption">Figure from Keasling’s research: Sulfide production by phsABC in various plasmids. pSB74 (orange) showed the highest reactivity.</div> | <div id="caption">Figure from Keasling’s research: Sulfide production by phsABC in various plasmids. pSB74 (orange) showed the highest reactivity.</div> | ||
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+ | (2)Promoter</b> <p>Promoter used was designed by Caitlin Conboy and was found within the parts registry. This promoter is a Quad Part Inverter: “that is, a PoPS-based inverter composed of four sub-parts: a ribosome binding site, a coding region for a repressor protein (e.g., lambda cI), a terminator, and the promoter (e.g., pLambda) regulated by the encoded repressor protein.” Research into promoter activity by previous groups has suggested that this promoter has a strong on state with a noticeable background in the off state .</p> | ||
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+ | (3) Promoter B0034</b> | ||
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+ | Biobrick Part:BBa_Q04121. | ||
+ | Length 1370 bp | ||
+ | IPTG-induced (regulatory) </p> | ||
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<a id="link" href="javascript:ReverseDisplay('background')">Read more about the background of the PHS gene</a> | <a id="link" href="javascript:ReverseDisplay('background')">Read more about the background of the PHS gene</a> |
Revision as of 01:03, 28 October 2010
our project
experimental methods
Our plasmid is composed of three parts: a promoter and a terminator Biobrick as well as a novel addition to the biobrick library, the phsABC gene that is known to encode Thiosulfate Reductase.
(1) phsABC gene and vectorThis central component encodes Thiosulfate Reductase. The gene phsABC was obtained from Dr. Jay Keasling laboratory at University of California, Berkeley. According to their results, Thiosulfate Reductase encoded in the plasmid pSB74 showed the highest activity, so we obtained phsABC from the plasmid pSB74. E. coli DH5α strain were used for plasmid manipulation.
Promoter used was designed by Caitlin Conboy and was found within the parts registry. This promoter is a Quad Part Inverter: “that is, a PoPS-based inverter composed of four sub-parts: a ribosome binding site, a coding region for a repressor protein (e.g., lambda cI), a terminator, and the promoter (e.g., pLambda) regulated by the encoded repressor protein.” Research into promoter activity by previous groups has suggested that this promoter has a strong on state with a noticeable background in the off state .
(3) Promoter B0034Biobrick Part:BBa_Q04121. Length 1370 bp IPTG-induced (regulatory)
Read more about the background of the PHS gene