Difference between revisions of "Part:BBa K4387987"

Revision as of 10:54, 6 October 2022

Hemolysin A secretion system for E. coli

The hemolysin A secretion machinery is a one-step secretion system (T1SS), originally isolated from uropathogenic E. coli strains. [1] It comprises three main peptides, the inner membrane proteins HlyB and HlyD, and the outer membrane protein TolC. Together, these three proteins build a continuous channel through which originally the HlyA toxin is secreted in a one-step manner. Interestingly, the secretion signal is not found on the N-terminal site, instead it is found at the C-terminal end and the signal sequence is not removed during secretion. Scientists have identified the secretion signal and were able to secrete various proteins of different sizes with this secretion machinery. [1]

This composite part contains the 2 membrane proteins HlyB (BBa_K4387999) and HlyD (BBa_K4387998) necessary for the channel formation. TolC is not included because it is already genomically expressed in many E. coli strains. [1] The formation of the secretion machinery is regulated by the constitutive promoter BBa_J23100 together with the RBS BBa_B0030. For a successful secretion, the c-terminal HlyA-tag (BBa_K4387997) has to be fused to the end of the protein intended to be secreted.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 7
Illegal NheI site found at 30
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1160
Illegal AgeI site found at 1330
1000
COMPATIBLE WITH RFC[1000]

References

[1] Ruano-Gallego, D., Fraile, S., Gutierrez, C. et al. Screening and purification of nanobodies from E. coli culture supernatants using the hemolysin secretion system. Microb Cell Fact 18, 47 (2019). https://doi.org/10.1186/s12934-019-1094-0

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K4387987 short</partinfo>
-The hemolysin A secretion machinery is a one-step secretion system (T1SS) that was originally isolated from uropathogenic E. coli strains. It is composed of 3 main peptides, the inner membrane proteins HlyB and HlyD, and the outer membrane protein TolC. Together, these 3 proteins build a continuous channel, through which originally the HlyA toxin is secreted in a one-step manner. Interestingly, the secretion signal is not found on the N-terminal site, instead it is found at the C-terminal end and the signal sequence is not removed during secretion. Scientists have identified the secretion signal and by fusing it to different proteins, were able to secrete various proteins with this secretion machinery. This composite part containes the 2 membrane proteins HlyB and HlyD necessary for the channel formation. TolC is not included because it is already genomically expressed in many E. coli strains. [1] The formation of the secretion machinery is regulated by the constitutive promoter <partinfo>J23100</partinfo> together with the RBS <partinfo>BBa_B0030</partinfo>.
+The hemolysin A secretion machinery is a one-step secretion system (T1SS), originally isolated from uropathogenic E. coli strains. [1] It comprises three main peptides, the inner membrane proteins HlyB and HlyD, and the outer membrane protein TolC. Together, these three proteins build a continuous channel through which originally the HlyA toxin is secreted in a one-step manner. Interestingly, the secretion signal is not found on the N-terminal site, instead it is found at the C-terminal end and the signal sequence is not removed during secretion. Scientists have identified the secretion signal and were able to secrete various proteins of different sizes with this secretion machinery. [1]
+This composite part contains the 2 membrane proteins HlyB (<partinfo>BBa_K4387999</partinfo>) and HlyD (<partinfo>BBa_K4387998</partinfo>) necessary for the channel formation. TolC is not included because it is already genomically expressed in many E. coli strains. [1] The formation of the secretion machinery is regulated by the constitutive promoter <partinfo>BBa_J23100</partinfo> together with the RBS <partinfo>BBa_B0030</partinfo>. For a successful secretion, the c-terminal HlyA-tag (<partinfo>BBa_K4387997</partinfo>) has to be fused to the end of the protein intended to be secreted.
 <!-- Add more about the biology of this part here