Difference between revisions of "Part:BBa K5323892"

Latest revision as of 15:13, 1 October 2024

P_tac-RBS-golS-P_gol-RBS-cI-TT-P_R

Uses a factor strong RBS(BBa_J34801), golS (BBa_K4468005), P_gol(BBa_K4468001), cI (BBa_R1051), P_tac(BBa_K180000) , TT(BBa_B0015) and PR(BBa_K2229450). This part is an easy BioBrick.

Usage and Biology

This component can determine whether the downstream gene is expressed by specific test gold.The cI expression product in this component represses its downstream promoter P_R in this component. When cI is inhibited, inhibition of the promoter P_R is lifted. When gold was detected, the downstream gene was not expressed, and when gold was not present, the downstream gene was expressed. The kill stwich designed in our project ensures that engineered bacteria can only survive in an environment containing Au[Ⅲ]. When Au[Ⅲ] is not present, the P_R promoter is not inhibited, initiating transcription, and mazF expresses the toxic protein MazF to kill the engineered bacteria. Once Au[Ⅲ] is present, the promoter P_R is inhibited, mazF does not express MazF toxic protein, and the engineered bacteria survive. We successfully verified this result by using E.coli BL21 as the chassis cell.

Fig 1 The growth curves of E.coli BL21. I + A is in the presence of the inducer IPTG and Au[Ⅲ]. IPTG is the only inducer, and Au[Ⅲ] is the only Au[Ⅲ]. The control group is without IPTG and Au[Ⅲ]

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1635
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 3721
Illegal AgeI site found at 1969
Illegal AgeI site found at 2318
Illegal AgeI site found at 3256
1000
COMPATIBLE WITH RFC[1000]

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 Uses a factor strong RBS(BBa_J34801), <i>golS</i> (BBa_K4468005), <i>P<sub>gol</sub></i>(BBa_K4468001), <i>cI</i> (BBa_R1051), <i>P<sub>tac</sub></i>(BBa_K180000) , TT(BBa_B0015) and <i>PR</i>(BBa_K2229450). This part is an easy BioBrick.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+This component can determine whether the downstream gene is expressed by specific test gold.The <i>cI</i> expression product in this component represses its downstream promoter <i>P<sub>R</sub></i> in this component. When <i>cI</i> is inhibited, inhibition of the promoter <i>P<sub>R</sub></i> is lifted. When gold was detected, the downstream gene was not expressed, and when gold was not present, the downstream gene was expressed.
+The kill stwich designed in our project ensures that engineered bacteria can only survive in an environment containing Au[Ⅲ]. When Au[Ⅲ] is not present, the <i>P<sub>R</sub></i> promoter is not inhibited, initiating transcription, and <i>mazF</i> expresses the toxic protein MazF to kill the engineered bacteria. Once Au[Ⅲ] is present, the promoter <i>P<sub>R</sub></i> is inhibited, <i>mazF</i> does not express MazF toxic protein, and the engineered bacteria survive. We successfully verified this result by using <i>E.coli</i> BL21 as the chassis cell.
+<center>https://static.igem.wiki/teams/5323/parts/j5.png</center>
+<center>Fig 1 The growth curves of <i>E.coli</i> BL21. I + A is in the presence of the inducer IPTG and Au[Ⅲ]. IPTG is the only inducer, and Au[Ⅲ] is the only Au[Ⅲ]. The control group is without IPTG and Au[Ⅲ]</center>
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