Difference between revisions of "Part:BBa K4340609"

Revision as of 03:21, 11 October 2022

genetic pH shooting system

To improve glsA (BBa_K4340611) and ldhA (BBa_K4340613) and to design a plasmid that is suitable for the hydroponics system, we designed our genetic pH shooting system.

genetic pH shooting system (BBa_K4340609) and pET11a empty vector pH maintenance functional test

Experiment 1: pH change and OD value

        
       
       Figure 1. The pH change of genetic pH shooting system_pET11a in the pH 5 initial environment
       Figure 2. The pH change of genetic pH shooting system_pET11a in the pH 6 initial environment.

       
       Figure 3. The pH change of genetic pH shooting system_pET11a in the pH 7 initial environment.

       
       Figure 4. The pH change of genetic pH shooting system_pET11a in the pH 8 initial environment.

       
       Figure 5. The pH change of genetic pH shooting system_pET11a in the pH 9 initial environment.

The pH change of the genetic pH shooting system is larger than the control group (pET11a) in the initial pH 5 environment in the first 5 hours, indicating that the genetic pH shooting system worked to converge the pH to neutral pH level. (Figure 1)

In the initial pH 6 environment, the convergence of the genetic pH shooting system to neutral pH performed well in 7th to 9th hours. In the following 15 hours, both the pH levels of the control and genetic pH shooting system group raised to pH 8 due to the possibility of the ammonia generated by the died E.coli. (Figure 2)

In the initial pH 7 environment, the pH curve of both groups are relatively similar, showing that the system does not function in a pH 7 environment, which conforms to the promoter design (Pasr for acidic environment and P-atp2 for alkaline environment) (Figure 3)

In both initial pH 8 and pH 9, the pH level of the genetic pH shooting system drops more than the control group (pET11a). This demonstrated that the base shooting circuit functioned to neutralize the alkaline environment. (Figure 4 & 5)

       
       
       Figure 5. The OD change of genetic pH shooting system_pET11a in the pH 5 initial environment.
       Figure 6. The OD change of genetic pH shooting system_pET11a in the pH 6 initial environment.

       
       Figure 7. The OD change of genetic pH shooting system_pET11a in the pH 7 initial environment.

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For the OD change of the genetic pH shooting system (Figure 7), The highest OD value is in the pH 7 environment, followed by pH 8, pH 6, pH 5, and pH 9. The OD value of pH 9 is clearly lower than the other pH groups (pH 5, 6, 7, 8, and 9 of genetic pH shooting system), which shows that the transforemd E.coli might not grow as well as E.coli with empty pET11a vector since it has to produce alkaline.

Experiment 2: Western blot

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