Team:PKU Beijing/Project/AND Gate 1/AND Gate Result

AND Gate of Shan Shen

This AND Gate is made of luxP-supD & Plac-T7ptag. The typical pattern of this AND gate is like this: (picture from K228119)

This family is the first AND gate family constructed by us, it had made us much more confident about other AND gates which were under construction at that time. This AND gates family contents nine AND gates with the difference on the RBS of the T7ptag: K228119, K228120, K228121, K228122, K228123, K228124, K228125, K228126, and K228127. And these gates are on the plasmid pSB4K5.

In order to test the function of these AND gates, Shan Shen 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, she transformed the AND gate plasmids into it, and tested them using various combinations of different concentration of IPTG and HSL (homoserine lactone). The result is tested by a microplate reader (Fig 1, 2, 3 and 4. For more detail, refer to our protocol)

Fig1. GFP expression of AND gate, K228125: lux-SupD-lac-T7ptag (rbs: J44001). The data are showed for (left to right): 1×10^-9, 1×10^-8, 1×10^-7, 1×10^-6, 1×10^-5 1×10^-4 and 1×10^-3M arabinose, and (bottom to top) 1×10^-9, 1×10^-8, 1×10^-7, 1×10^-6, 1×10^-5 and 1×10^-4M HSL.

Fig2. GFP expression of AND gate, K228124: lux-SupD-lac-T7ptag (rbs: J61127). The data are showed for (left to right): 1×10^-9, 1×10^-8, 1×10^-7, 1×10^-6, 1×10^-5 1×10^-4 and 1×10^-3M arabinose, and (bottom to top) 1×10^-9, 1×10^-8, 1×10^-7, 1×10^-6, 1×10^-5 and 1×10^-4M HSL.

Fig3. GFP expression of AND gate, K228120: lux-SupD-lac-T7ptag (rbs: B0032). The data are showed for (left to right): 1×10^-9, 1×10^-8, 1×10^-7, 1×10^-6, 1×10^-5 1×10^-4 and 1×10^-3M arabinose, and (bottom to top) 1×10^-8, 1×10^-7, 1×10^-6, 1×10^-5 and 1×10^-4M HSL.

Fig4. GFP expression of AND gate, K228126 : lux-SupD-lac-T7ptag (rbs: B0033). The data are showed for (left to right): 1×10^-9, 1×10^-8, 1×10^-7, 1×10^-6, 1×10^-5 1×10^-4 and 1×10^-3M arabinose, and (bottom to top) 1×10^-9, 1×10^-8, 1×10^-7, 1×10^-6, 1×10^-5 and 1×10^-4M HSL.

From the result of some of these AND gates, we found that many of these AND gates are inducible by IPTG, but independent of the concentration of HSL. It is obvious that the leakage of lux system is significant, and the basal expression of SupD is so high, that without HSL there is sufficient SupD to work on T7ptag. This result is consistent with the sensor result of lux system: there are quite amount of expression without inducing of HSL (for more detail, refer to Sensor Result).

The result of other five AND gates are not shown here.

AND Gate of Shuke Wu

AND Gate of Guosheng Zhang (The Best)

Other AND Gates

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