Difference between revisions of "Part:BBa K2922025"

Revision as of 03:02, 20 October 2019

Colicin-E1 immunity protein coding region

Summary

Abbreviated as Eimm, the molecular weight of Colicin-E1 immunity protein is 13 kDa, and the protein is encoded by gene imm. In wild strains of E.coli, the protein is able to protect cells, which harbors the plasmid ColE1 encoding Colicin-E1, against Colicin-E1. Immunity proteins interact with the pore-forming domain in the inner membrane. And the inactivation of Colicin-E1 occurs via interactions between the voltage-gated region and the transmembrane helices of the immunity protein. This protein will prevent E.coli secreting Colicin-E1 from killing themselves.[1] (Fig.1)

Fig.1 Eimm prevents E.coli BL21 (DE3) secreting Colicin-E1 from killing themselves.

Identification

When we were building this circuit, we did the nucleic acid gel electrophoresis experiment to verify. After the circuit was built, we sent the plasmid to sequence, and got the correct sequencing. After new molecular cloning experiments, we did Enzyme-Cut identification to certify the plasmid was correct. We used the XbaI and PstI to cut the plasmid, then we got the target separate fragment- 342bp. (Fig.2)

Fig.2 The result of this plasmid cut with enzyme XbaI and PstI.

Reference

[1] H. Y. Song, W. A. Cramer, Membrane topography of ColE1 gene products: the immunity protein. Journal of Bacteriology 173, 2935 (1991).

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 4: / Line 4: @@
 ===Summary===
 Abbreviated as Eimm, the molecular weight of Colicin-E1 immunity protein is 13 kDa, and the protein is encoded by gene <i>imm</i>. In wild strains of <i>E.coli</i>, the protein is able to protect cells, which harbors the plasmid ColE1 encoding Colicin-E1, against Colicin-E1. Immunity proteins interact with the pore-forming domain in the inner membrane. And the inactivation of Colicin-E1 occurs via interactions between the voltage-gated region and the transmembrane helices of the immunity protein. This protein will prevent <i>E.coli</i> secreting Colicin-E1 from killing themselves.[1] (Fig.1)
-<table><tr><th>[[File:E1design.png|thumb|300px|Fig.1 Eimm prevents <i>E.coli</i> BL21 (DE3) secreting Colicin-E1 from killing themselves.]]</th><th></table>
+<table><tr><th>[[File:E1design.png|thumb|720px|Fig.1 Eimm prevents <i>E.coli</i> BL21 (DE3) secreting Colicin-E1 from killing themselves.]]</th><th></table>
 ===Identification===
-When we are building this circuit, we are doing the nucleic acid gel electrophoresis experiment to verify. After the circuit is built, we send the plasmid to sequence, and get the correct sequencing.
+When we were building this circuit, we did the nucleic acid gel electrophoresis experiment to verify. After the circuit was built, we sent the plasmid to sequence, and got the correct sequencing.
-After new molecular cloning experiments, we do Enzyme-Cut identification to certify the plasmid is correct. We use the <i>Xba</i>I and <i>Pst</i>I to cut the plasmid, then we get the target separate fragment- 342bp. (Fig.2)
+After new molecular cloning experiments, we did Enzyme-Cut identification to certify the plasmid was correct. We used the <i>Xba</i>I and <i>Pst</i>I to cut the plasmid, then we got the target separate fragment- 342bp. (Fig.2)
 <table><tr><th>[[File:Eimm Nimm.png|thumb|300px|Fig.2 The result of this plasmid cut with enzyme <i>Xba</i>I and <i>Pst</i>I.]]</th><th></table>
 ===Reference===