Team:PKU Beijing/Project/AND Gate 1/AND Gate Result
From 2009.igem.org
(→AND Gates of Shan Shen) |
(→AND Gates of Shuke Wu) |
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This family is the AND gate family constructed also the quickliest: at the same time with Shan Shen's AND gates. | This family is the AND gate family constructed also the quickliest: at the same time with Shan Shen's AND gates. | ||
- | + | Nine AND gates sharing the same promoters and with the difference on the RBS of the T7ptag are in this family: K228823, K228824, K228825, K228826, K228827, K228828, K228829, K228830, and K228831. And these gates are also on the plasmid pSB4K5. | |
In order to test the function of these AND gates, Shuke use a reporter GFP under the control of a T7 promoter: K228099. And the reporter is on the plasmid J61002. | In order to test the function of these AND gates, Shuke use a reporter GFP under the control of a T7 promoter: K228099. And the reporter is on the plasmid J61002. | ||
- | Using the E.coli with the reporter plasmid, he transformed the AND gate plasmids into it, and tested them using various combinations of different | + | Using the E.coli with the reporter plasmid, he transformed the AND gate plasmids into it, and tested them using various combinations of different concentrations of IPTG and arabinose. The result is tested by a microplate reader (Fig 1, 2, 3 and 4. For more detail, refer to our [[Team:PKU_Beijing/Notebook/Protocol/Chemical_Inducible_GFP|protocol]]) |
[[Image:PKU_WSK_2K.png|600px|center|thumb|Fig1. GFP expression of AND gate, K228829: lac-SupD-pBad-T7ptag(rbs: B0033). The data are showed for (left to right): 1×10^-9, 1×10^-7, 1×10^-6, 1×10^-5 1×10^-4 and 1×10^-3M IPTG, and (bottom to top) 1×10^-9, 1×10^-7, 1×10^-6, 1×10^-5, 1×10^-4 and 1×10^-3M arabinose.]] | [[Image:PKU_WSK_2K.png|600px|center|thumb|Fig1. GFP expression of AND gate, K228829: lac-SupD-pBad-T7ptag(rbs: B0033). The data are showed for (left to right): 1×10^-9, 1×10^-7, 1×10^-6, 1×10^-5 1×10^-4 and 1×10^-3M IPTG, and (bottom to top) 1×10^-9, 1×10^-7, 1×10^-6, 1×10^-5, 1×10^-4 and 1×10^-3M arabinose.]] | ||
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[[Image:PKU_WSK_1J_4.png|200px|left|thumb|Fig4. GFP expression of AND gate, K228823: simplified version of Fig3. The data are showed for 1×10^-9 and 1×10^-2M IPTG, and 1×10^-9 and 1×10^-2M arabinose. The concentration of 1×10^-9 equals input=0, and 1×10^-2 equals input=1. So the four columns show the response of (0,0), (0,1), (1,0) and (1,1).]] | [[Image:PKU_WSK_1J_4.png|200px|left|thumb|Fig4. GFP expression of AND gate, K228823: simplified version of Fig3. The data are showed for 1×10^-9 and 1×10^-2M IPTG, and 1×10^-9 and 1×10^-2M arabinose. The concentration of 1×10^-9 equals input=0, and 1×10^-2 equals input=1. So the four columns show the response of (0,0), (0,1), (1,0) and (1,1).]] | ||
- | Discussion: AND gate K228829 (Fig 1) and K228823 (Fig 3) are the two best AND gates in this family, while others did not display AND gate behavior (Fig 2). It is a common sense that the activity range of promoters affects the function of the AND gate circuit (Anderson JC, Voigt CA, Arkin AP (2007) Environmental signal integration by a modular AND gate. Mol Syst Biol 3: 133). Thus we used nine different RBSs as a method to modulate the expression level of T7ptag and got some quite good AND gates (Fig 1&3). However, this strategy can not change the transcription of SupD, which may lead to leakage of AND gates. Because Shuke used lac system, which suffers from some problems (refer to our sensor result), to drive the SupD, this AND gate family can not have some perfect AND gates. | + | Discussion: AND gate K228829 (Fig 1) and K228823 (Fig 3) are the two best AND gates in this family, while others did not display AND gate behavior (Fig 2). It is a common sense that the activity range of promoters affects the function of the AND gate circuit (Anderson JC, Voigt CA, Arkin AP (2007) Environmental signal integration by a modular AND gate. Mol Syst Biol 3: 133). Thus we used nine different RBSs as a method to modulate the expression level of T7ptag and got some quite good AND gates (Fig 1&3). However, this strategy can not change the transcription level of SupD, which may lead to leakage of AND gates. Because Shuke used lac system, which suffers from some problems (refer to our sensor result), to drive the SupD, this AND gate family can not have some perfect AND gates. |
- | To improve, we should choose a | + | To improve, we should choose a tight promoter (pSal or pBad), with little leakage, to drive SupD. |
- | The | + | The results of other six AND gates are not shown here. |
==='''AND Gate of Guosheng Zhang (The Best AND Gate we achieve)'''=== | ==='''AND Gate of Guosheng Zhang (The Best AND Gate we achieve)'''=== |
Revision as of 09:26, 20 October 2009
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