Difference between revisions of "Part:BBa K245114:Experience"
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= Colony Result =
 
= Colony Result =
−
We successfully built DenTeeth with LL-37 .
+
  We successfully built DenTeeth with LL-37 .
 
[[File:T--NCTU_Formosa--seq_abc_pcr.jpg--seq_abc_pcr.jpg|300px|thumb|center|'''Figure 1. Colony PCR result of DenTeeth after colony into E. coli BL21. (4685 b.p.)
 
[[File:T--NCTU_Formosa--seq_abc_pcr.jpg--seq_abc_pcr.jpg|300px|thumb|center|'''Figure 1. Colony PCR result of DenTeeth after colony into E. coli BL21. (4685 b.p.)
 
]]
 
]]
  
 
= Model =
 
= Model =
−
Through the modeling built with substituting parameters from published articles, the growth of E. coli and P. gingivalis under the effect of LL-37 would be inhibited significantly. However, the figure also shows that E. coli could still live with LL-37. Simply speaking, we successfully test the feasibility of DenTeeth and find that we can improve the DenTeeth with both biobrick design and efficiency optimization model.
+
  Through the modeling built with substituting parameters from published articles, the growth of E. coli and P. gingivalis under the effect of LL-37 would be inhibited significantly. However, the figure also shows that E. coli could still live with LL-37. Simply speaking, we successfully test the feasibility of DenTeeth and find that we can improve the DenTeeth with both biobrick design and efficiency optimization model.
 
[[File:T--NCTU_Formosa--bacteria_V.S._time.png|500px|thumb|center|'''Figure 2. The growth curve of E. coli and P.gingivalis
 
[[File:T--NCTU_Formosa--bacteria_V.S._time.png|500px|thumb|center|'''Figure 2. The growth curve of E. coli and P.gingivalis
 
]]
 
]]
 
[[File:T--NCTU_Formosa--bacteria_with_DenTeeth.png|500px|thumb|center|'''Figure 3. The growth curve of E. coli and P.gingivalis
 
[[File:T--NCTU_Formosa--bacteria_with_DenTeeth.png|500px|thumb|center|'''Figure 3. The growth curve of E. coli and P.gingivalis
 
]]
 
]]
−
 Because P. gingivalis was in the RG2, so we could not do the LL-37 functional test by inhibit the growth of P. gingivalis. After reading some related papers, we found that the killing rate of E. coli was similar to that of P. gingivalis [4]. Based on these data, we determined to make E. coli, DH5α with pET32A, as the bacteria killed by DenTeeth in the LL-37 functional test.
+
  Because P. gingivalis was in the RG2, so we could not do the LL-37 functional test by inhibit the growth of P. gingivalis. After reading some related papers, we found that the killing rate of E. coli was similar to that of P. gingivalis [4]. Based on these data, we determined to make E. coli, DH5α with pET32A, as the bacteria killed by DenTeeth in the LL-37 functional test.
 
[[File:T--NCTU_Formosa--bacteriatable.png|500px|thumb|center|'''Table 1. Parameters of the sterilization system of LL37
 
[[File:T--NCTU_Formosa--bacteriatable.png|500px|thumb|center|'''Table 1. Parameters of the sterilization system of LL37
 
]]
 
]]
  
 
= Functional Test =
 
= Functional Test =
−
After finishing the design of DenTeeth, we wanted to know whether its inhibition ability can have a function, so we designed the following experiment.
+
  After finishing the design of DenTeeth, we wanted to know whether its inhibition ability can have a function, so we designed the following experiment.
−
By taking the method of dish culture and sticking up the filter paper, we dropped the DenTeeth on the filter in the center of the E. coli, DH5α with pET32A, plate. After twelve hours, a circular area around the spot of the DenTeeth formed, in which the bacteria colonies did not grow. Zone of inhibition proved that the LL-37 produced by DenTeeth could successfully secrete out and inhibit the growth of E. coli, DH5α with pET32A.
+
  By taking the method of dish culture and sticking up the filter paper, we dropped the DenTeeth on the filter in the center of the E. coli, DH5α with pET32A, plate. After twelve hours, a circular area around the spot of the DenTeeth formed, in which the bacteria colonies did not grow. Zone of inhibition proved that the LL-37 produced by DenTeeth could successfully secrete out and inhibit the growth of E. coli, DH5α with pET32A.
 
[[File:T--NCTU_Formosa--Zone of inhibition.png|500px|thumb|center|'''Figure 4. LL-37 Zone of inhibition
 
[[File:T--NCTU_Formosa--Zone of inhibition.png|500px|thumb|center|'''Figure 4. LL-37 Zone of inhibition
 
]]
 
]]

Revision as of 17:28, 21 October 2021


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K245114

User Reviews

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iGEM NCTU_Formosa(2021)

Colony Result

  We successfully built DenTeeth with LL-37 .

Figure 1. Colony PCR result of DenTeeth after colony into E. coli BL21. (4685 b.p.)

Model

  Through the modeling built with substituting parameters from published articles, the growth of E. coli and P. gingivalis under the effect of LL-37 would be inhibited significantly. However, the figure also shows that E. coli could still live with LL-37. Simply speaking, we successfully test the feasibility of DenTeeth and find that we can improve the DenTeeth with both biobrick design and efficiency optimization model.

Figure 2. The growth curve of E. coli and P.gingivalis
Figure 3. The growth curve of E. coli and P.gingivalis

  Because P. gingivalis was in the RG2, so we could not do the LL-37 functional test by inhibit the growth of P. gingivalis. After reading some related papers, we found that the killing rate of E. coli was similar to that of P. gingivalis [4]. Based on these data, we determined to make E. coli, DH5α with pET32A, as the bacteria killed by DenTeeth in the LL-37 functional test.

Table 1. Parameters of the sterilization system of LL37

Functional Test

  After finishing the design of DenTeeth, we wanted to know whether its inhibition ability can have a function, so we designed the following experiment.   By taking the method of dish culture and sticking up the filter paper, we dropped the DenTeeth on the filter in the center of the E. coli, DH5α with pET32A, plate. After twelve hours, a circular area around the spot of the DenTeeth formed, in which the bacteria colonies did not grow. Zone of inhibition proved that the LL-37 produced by DenTeeth could successfully secrete out and inhibit the growth of E. coli, DH5α with pET32A.

Figure 4. LL-37 Zone of inhibition
;

BBa_K245114 NCTU-Formosa(2021) Not understood UNIQ62a93c62b5e6e0b2-partinfo-00000003-QINU