Difference between revisions of "Part:BBa K4593020"
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This part contains 3 genetic circuits: an S. aureus quorum sensing (QS) detection system (BBa_I746101), an Endolysin combinative expression system (BBa_K4593019), and a self-lysing release system (BBa_I746104 and BBa_K4593001). | This part contains 3 genetic circuits: an S. aureus quorum sensing (QS) detection system (BBa_I746101), an Endolysin combinative expression system (BBa_K4593019), and a self-lysing release system (BBa_I746104 and BBa_K4593001). | ||
| − | As the final intended product of our project's S. aureus elimination module, this device could constitutively express S. aureus targeting endolysins (a set of enzymes that specifically lyses S. aureus) and detect the presence of S. aureus QS signal. If the QS signal molecule is detected, the self-lysing enzyme composite (Spn1s_LysRZ) will be activated, and the engineered cell will lyse itself to release the endolysin that eliminates pathogenic S. aureus. | + | As the final intended product of our project's S. aureus elimination module, this device could constitutively express S. aureus targeting endolysins (a set of enzymes that specifically lyses S. aureus) and detect the presence of S. aureus QS signal. If the QS signal molecule is detected, the self-lysing enzyme composite (Spn1s_LysRZ) will be activated, and the engineered cell will lyse itself to release the endolysin that eliminates pathogenic S. aureus. |
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| + | <html> | ||
| + | <figure> | ||
| + | <p style="text-align:center;"><img src="https://static.igem.wiki/teams/4593/wiki/design/design-15.png" width="700" height="auto"/> | ||
| + | </figcaption> | ||
| + | </figure> | ||
| + | </html> | ||
| + | Fig.1 An abbreviated demonstration of the single cell gene circuit of S. aureus in vivo elimination device. Pro 1 and Pro 3 are both constitutive promoters. RBS and terminator are omitted for the sake of space. | ||
===Endolysin expression circuit=== | ===Endolysin expression circuit=== | ||
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===QS-detection circuit=== | ===QS-detection circuit=== | ||
| − | This circuit could constitutively express AgrA and AgrC, two proteins that transduce the QS signal. Specifically, AgrC is a membrane that recognizes extracellular AIPs (QS molecule of S. aureus). After AgrC detects the QS signal, AgrA will be subsequently phosphorylated by it, gaining the ability to activate the transcription of genes downstream promoter P2. | + | This circuit could constitutively express AgrA and AgrC, two proteins that transduce the QS signal. Specifically, AgrC is a membrane kinase that recognizes extracellular AIPs (QS molecule of S. aureus). After AgrC detects the QS signal, AgrA will be subsequently phosphorylated by it, gaining the ability to activate the transcription of genes downstream promoter P2. |
===Release circuit=== | ===Release circuit=== | ||
Latest revision as of 03:39, 11 October 2023
S. aureus in vivo elimination apparatus for E. coli
Overview
This part contains 3 genetic circuits: an S. aureus quorum sensing (QS) detection system (BBa_I746101), an Endolysin combinative expression system (BBa_K4593019), and a self-lysing release system (BBa_I746104 and BBa_K4593001).
As the final intended product of our project's S. aureus elimination module, this device could constitutively express S. aureus targeting endolysins (a set of enzymes that specifically lyses S. aureus) and detect the presence of S. aureus QS signal. If the QS signal molecule is detected, the self-lysing enzyme composite (Spn1s_LysRZ) will be activated, and the engineered cell will lyse itself to release the endolysin that eliminates pathogenic S. aureus.
Endolysin expression circuit
This circuit could constitutively express 3 types of S. aureus targeting endolysin-LysDZ25, ClyC, and Lys GH15-that have maximum efficiency conditions complementary with each other. This ensures that at least one type of endolysin will be functional under various conditions in the human digestive tract. The lytic efficiency of the endolysins with respect to concentration is being characterized separately in our project. For detailed information, see the following basic parts:
LysDZ25: BBa_K4593000
ClyC: BBa_K4593002
LysGH25: BBa_K4593003
QS-detection circuit
This circuit could constitutively express AgrA and AgrC, two proteins that transduce the QS signal. Specifically, AgrC is a membrane kinase that recognizes extracellular AIPs (QS molecule of S. aureus). After AgrC detects the QS signal, AgrA will be subsequently phosphorylated by it, gaining the ability to activate the transcription of genes downstream promoter P2.
Release circuit
The circuit allows the engineered bacteria to express the self-lysing enzyme complex (Spn1s_LysRZ) under the presence of S. aureus QS signal. Specifically, phosphorylated AgrA could activate the transcription activity of the P2 promoter, enabling the Spn1s_LysRZ located downstream to be transcribed.
For detailed characterization of Spn1s_LysRZ, see BBa_K4593001
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal NheI site found at 3468
Illegal NheI site found at 3491 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 418
Illegal BglII site found at 6040
Illegal BamHI site found at 1302
Illegal XhoI site found at 2591 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 5684
Illegal NgoMIV site found at 6524
Illegal NgoMIV site found at 7198
Illegal AgeI site found at 3090
Illegal AgeI site found at 3228
Illegal AgeI site found at 5553
Illegal AgeI site found at 6668
Illegal AgeI site found at 6896
Illegal AgeI site found at 7067
Illegal AgeI site found at 7180 - 1000COMPATIBLE WITH RFC[1000]