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+ | <div class="sub-content"> | ||
+ | <div class="sub-title">Parts Contribution</div> | ||
+ | <div class="article-title">BBa_K4028000</div> | ||
+ | <div class="article-content"><b>== Profile ==</b></div> | ||
+ | <div class="article-content"> | ||
+ | Name: tke2<br /> | ||
+ | Base Pairs: 4158 bp<br /> | ||
+ | Origin: Pseudomonas putida KT2440, genome<br /> | ||
+ | Properties: Anti-bacterial effector in type VI secretion system | ||
+ | </div> | ||
+ | <div class="article-content"><b>== Usage and Biology ==</b></div> | ||
+ | <div class="article-content">BBa_K4028000 is a coding sequence of tke2 from Pseudomonas putida KT2440. Tke2, | ||
+ | known as a toxic Rhs-type effector, is an effect factor in type VI secretion system (T6SS). | ||
+ | A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs, including opportunistic | ||
+ | pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit the human gut | ||
+ | (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6] Under natural | ||
+ | conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or antibacterial effects | ||
+ | (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical contact, thus inhibiting | ||
+ | the growth of recipient cells. The bacteria encoding T6SS can translate and produce corresponding immune | ||
+ | proteins to counteract the damage caused by toxic effector factors.[1,2,3] | ||
+ | </div> | ||
+ | <div class="img-wrap"> | ||
+ | <img src="https://static.igem.org/mediawiki/2021/0/01/T--Shanghai_Metro--contribution01.jpg" alt=""> | ||
+ | <span>Figure1. Principle diagram of T6SS.</span> | ||
+ | </div> | ||
+ | <div class="article-title">BBa_K4028001</div> | ||
+ | <div class="article-content"><b>== Profile ==</b></div> | ||
+ | <div class="article-content"> | ||
+ | Name: ike2<br /> | ||
+ | Base Pairs: 447bp<br /> | ||
+ | Origin: Pseudomonas putida KT2440, genome<br /> | ||
+ | Properties: Immunity effector in type VI secretion system | ||
+ | </div> | ||
+ | <div class="article-content"><b>== Usage and Biology ==</b></div> | ||
+ | <div class="article-content"> | ||
+ | BBa_K4028001 is a coding sequence of ike2, an immunity protein in Pseudomonas putida KT2440. Ike2 is used | ||
+ | for protecting bacteria from type VI secretion system (T6SS). <br /> | ||
+ | A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs, including opportunistic | ||
+ | pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit the human gut | ||
+ | (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6] Under natural | ||
+ | conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or antibacterial effects | ||
+ | (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical contact, thus inhibiting | ||
+ | the growth of recipient cells. Meanwhile, the bacteria encoding T6SS can translate and produce corresponding | ||
+ | immune protein (ike2) to counteract the damage caused by toxic effector factors.[1,2,3] | ||
+ | </div> | ||
+ | <div class="article-title">BBa_K4028005</div> | ||
+ | <div class="article-content"><b>== Profile ==</b></div> | ||
+ | <div class="article-content"> | ||
+ | Name: tet promoter<br /> | ||
+ | Base Pairs: 56bp<br /> | ||
+ | Origin: Escherichia coli, synthetic<br /> | ||
+ | Properties: Inducible promoter regulated by tetracycline | ||
+ | </div> | ||
+ | <div class="article-content"><b>== Usage and Biology ==</b></div> | ||
+ | <div class="article-content"> | ||
+ | This is the naturally-occuring version of the TetR class B promoter. Note that this will promote | ||
+ | bidirectionally. (pR1 and pR2 will promote in the forward direction; pA will promote in the reverse | ||
+ | direction.) The three transcription start sites are not labeled, but can be found from the reference. <br /> | ||
+ | We set different tetracycline concentrations to test the protein expression induced by TetR(B) promoter. In | ||
+ | our design, the immune protein (ike4) is regulated by TetR(B) promoter. The expression of the immune protein | ||
+ | (ike4) directly affects and is positively correlated with strain growth. Therefore, the strain growth can | ||
+ | indicate the regulation of the TetR(B) promoter.<br /> | ||
+ | The results of the test are below. | ||
+ | </div> | ||
+ | <div class="img-wrap"> | ||
+ | <span>Table 1. OD600 value of the strain D: Pus232-ike4-tke4 after 6 hours</span> | ||
+ | <img src="https://static.igem.org/mediawiki/2021/c/cd/T--Shanghai_Metro--contribution02.jpg" alt=""> | ||
+ | </div> | ||
+ | <div class="img-wrap"> | ||
+ | <img src="https://static.igem.org/mediawiki/2021/2/27/T--Shanghai_Metro--contribution03.jpg" alt=""> | ||
+ | <span>Figure 1. Biomass Concentration curve of Pus323-tke4-ike4 Strain with tetracycline</span> | ||
+ | </div> | ||
+ | <div class="article-content">The results show that concentrations at the range of 300-500µg/L are more | ||
+ | effective. The results also proved that adding tetracycline to strain will boost the growth of strain. It | ||
+ | determines that the optimal concentration of tetracycline is 300µg/L to 500µg/L.</div> | ||
+ | <div class="article-title">BBa_K4028006</div> | ||
+ | <div class="article-content"><b>== Profile ==</b></div> | ||
+ | <div class="article-content"> | ||
+ | Name: tet pro-ike2<br /> | ||
+ | Base Pairs: 533 bp<br /> | ||
+ | Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic<br /> | ||
+ | Properties: Gene technology for protecting patented bacterial strains | ||
+ | </div> | ||
+ | <div class="article-content"><b>== Usage and Biology ==</b></div> | ||
+ | <div class="article-content">A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs, | ||
+ | including opportunistic pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit | ||
+ | the human gut (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6] | ||
+ | Under natural conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or | ||
+ | antibacterial effects (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical | ||
+ | contact, thus inhibiting the growth of recipient cells. Meanwhile, the bacteria encoding T6SS can translate | ||
+ | and produce corresponding immune protein to counteract the damage caused by toxic effector factors.[1,2,3] | ||
+ | </div> | ||
+ | <div class="article-content"><b>== Construct design ==</b></div> | ||
+ | <div class="article-content">The immunity effector ike2 is under tet promoter. The composite part is inserted in | ||
+ | the pUS232 vector (Figure 3).</div> | ||
+ | <div class="img-wrap"> | ||
+ | <img src="https://static.igem.org/mediawiki/2021/5/54/T--Shanghai_Metro--contribution04.jpg" alt=""> | ||
+ | <span>Figure 2. Immune protein expression box.</span> | ||
+ | </div> | ||
+ | <div class="img-wrap"> | ||
+ | <img src="https://static.igem.org/mediawiki/2021/c/c9/T--Shanghai_Metro--contribution05.jpg" alt=""> | ||
+ | <span>Figure 3. Schematic map of immune expression plasmids.</span> | ||
+ | </div> | ||
+ | <div class="article-title">BBa_K4028007</div> | ||
+ | <div class="article-content"><b>== Profile ==</b></div> | ||
+ | <div class="article-content"> | ||
+ | Name: tet pro-ike4<br /> | ||
+ | Base Pairs: 560 bp<br /> | ||
+ | Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic<br /> | ||
+ | Properties: Gene technology for protecting patented bacterial strains | ||
+ | </div> | ||
+ | <div class="article-content"><b>== Usage and Biology ==</b></div> | ||
+ | <div class="article-content">A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs, | ||
+ | including opportunistic pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit | ||
+ | the human gut (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6] | ||
+ | Under natural conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or | ||
+ | antibacterial effects (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical | ||
+ | contact, thus inhibiting the growth of recipient cells. Meanwhile, the bacteria encoding T6SS can translate | ||
+ | and produce corresponding immune protein to counteract the damage caused by toxic effector factors.[1,2,3] | ||
+ | </div> | ||
+ | <div class="article-content"><b>== Construct design ==</b></div> | ||
+ | <div class="article-content">The immunity effector ike4 is under tet promoter(Figure 2). This composite part is | ||
+ | inserted in the pUS232 vector (Figure 3).</div> | ||
+ | <div class="img-wrap"> | ||
+ | <img src="https://static.igem.org/mediawiki/2021/b/b9/T--Shanghai_Metro--contribution06.jpg" alt=""> | ||
+ | <span>Figure 2. Immune protein expression box.</span> | ||
+ | </div> | ||
+ | <div class="img-wrap"> | ||
+ | <img src="https://static.igem.org/mediawiki/2021/b/bc/T--Shanghai_Metro--contribution07.jpg" alt=""> | ||
+ | <span>Figure 3. Schematic map of immune expression plasmids.</span> | ||
+ | </div> | ||
+ | <div class="article-title">BBa_K4028008</div> | ||
+ | <div class="article-content"><b>== Profile ==</b></div> | ||
+ | <div class="article-content"> | ||
+ | Name: lac pro-tke2-tet pro-ike2<br /> | ||
+ | Base Pairs: 4746 bp<br /> | ||
+ | Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic<br /> | ||
+ | Properties: Gene technology for protecting patented bacterial strains | ||
+ | </div> | ||
+ | <div class="article-content"><b>== Usage and Biology ==</b></div> | ||
+ | <div class="article-content">A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs, | ||
+ | including opportunistic pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit | ||
+ | the human gut (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6] | ||
+ | Under natural conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or | ||
+ | antibacterial effects (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical | ||
+ | contact, thus inhibiting the growth of recipient cells. Meanwhile, the bacteria encoding T6SS can translate | ||
+ | and produce corresponding immune protein to counteract the damage caused by toxic effector factors.[1,2,3] | ||
+ | </div> | ||
+ | <div class="article-content"><b>== Construct design ==</b></div> | ||
+ | <div class="article-content">With this idea coming from the T6SS, we designed to express the effector components | ||
+ | of T6SS in industrial strains by inserting the gene segment tke2-ike2 to induce the controlled expression of | ||
+ | the corresponding immune proteins. The normal growth of industrial microorganisms can be ensured only by | ||
+ | adding certain components (tetracycline) in the fermentation broth, so as to achieve the purpose of the | ||
+ | experiment. </div> | ||
+ | <div class="img-wrap"> | ||
+ | <img src="https://static.igem.org/mediawiki/2021/7/7a/T--Shanghai_Metro--contribution08.jpg" alt=""> | ||
+ | <span>Figure2. The gene design of TetR promoter-ike2-lac promoter-tke2.</span> | ||
+ | </div> | ||
+ | <div class="img-wrap"> | ||
+ | <img src="https://static.igem.org/mediawiki/2021/2/27/T--Shanghai_Metro--contribution09.jpg" alt=""> | ||
+ | <span>Figure 3. Schematic map of T6SS effector and immune expression plasmids.</span> | ||
+ | </div> | ||
+ | <div class="article-title">BBa_K4028009</div> | ||
+ | <div class="article-content"><b>== Profile ==</b></div> | ||
+ | <div class="article-content"> | ||
+ | Name: lac pro-tke4-tet pro-ike4<br /> | ||
+ | Base Pairs: 5208 bp<br /> | ||
+ | Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic<br /> | ||
+ | Properties: Gene technology for protecting patented bacterial strains | ||
+ | </div> | ||
+ | <div class="article-content"><b>== Usage and Biology ==</b></div> | ||
+ | <div class="article-content">A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs, | ||
+ | including opportunistic pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit | ||
+ | the human gut (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6] | ||
+ | Under natural conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or | ||
+ | antibacterial effects (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical | ||
+ | contact, thus inhibiting the growth of recipient cells. Meanwhile, the bacteria encoding T6SS can translate | ||
+ | and produce corresponding immune protein to counteract the damage caused by toxic effector factors.[1,2,3] | ||
+ | </div> | ||
+ | <div class="article-content"><b>== Construct design ==</b></div> | ||
+ | <div class="article-content">Tke4 is a key functional factor that inhibits bacterial growth which is controlled | ||
+ | by a lac promoter (Figure 2). The immunity effector ike4 is under tet promoter. The anti-bacterial effector | ||
+ | tke4 and the immunity effector ike4 are inserted in the pUS232 vector (Figure 3).</div> | ||
+ | <div class="img-wrap"> | ||
+ | <img src="https://static.igem.org/mediawiki/2021/6/63/T--Shanghai_Metro--contribution10.jpg" alt=""> | ||
+ | <span>Figure 2. T6SS effector and immune protein expression box.</span> | ||
+ | </div> | ||
+ | <div class="img-wrap"> | ||
+ | <img src="https://static.igem.org/mediawiki/2021/a/ad/T--Shanghai_Metro--contribution11.jpg" alt=""> | ||
+ | <span>Figure 3. Schematic map of T6SS effector and immune expression plasmids.</span> | ||
+ | </div> | ||
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Latest revision as of 02:43, 20 October 2021
Parts Contribution
BBa_K4028000
== Profile ==
Name: tke2
Base Pairs: 4158 bp
Origin: Pseudomonas putida KT2440, genome
Properties: Anti-bacterial effector in type VI secretion system
Base Pairs: 4158 bp
Origin: Pseudomonas putida KT2440, genome
Properties: Anti-bacterial effector in type VI secretion system
== Usage and Biology ==
BBa_K4028000 is a coding sequence of tke2 from Pseudomonas putida KT2440. Tke2,
known as a toxic Rhs-type effector, is an effect factor in type VI secretion system (T6SS).
A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs, including opportunistic
pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit the human gut
(Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6] Under natural
conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or antibacterial effects
(amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical contact, thus inhibiting
the growth of recipient cells. The bacteria encoding T6SS can translate and produce corresponding immune
proteins to counteract the damage caused by toxic effector factors.[1,2,3]
Figure1. Principle diagram of T6SS.
BBa_K4028001
== Profile ==
Name: ike2
Base Pairs: 447bp
Origin: Pseudomonas putida KT2440, genome
Properties: Immunity effector in type VI secretion system
Base Pairs: 447bp
Origin: Pseudomonas putida KT2440, genome
Properties: Immunity effector in type VI secretion system
== Usage and Biology ==
BBa_K4028001 is a coding sequence of ike2, an immunity protein in Pseudomonas putida KT2440. Ike2 is used
for protecting bacteria from type VI secretion system (T6SS).
A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs, including opportunistic pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit the human gut (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6] Under natural conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or antibacterial effects (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical contact, thus inhibiting the growth of recipient cells. Meanwhile, the bacteria encoding T6SS can translate and produce corresponding immune protein (ike2) to counteract the damage caused by toxic effector factors.[1,2,3]
A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs, including opportunistic pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit the human gut (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6] Under natural conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or antibacterial effects (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical contact, thus inhibiting the growth of recipient cells. Meanwhile, the bacteria encoding T6SS can translate and produce corresponding immune protein (ike2) to counteract the damage caused by toxic effector factors.[1,2,3]
BBa_K4028005
== Profile ==
Name: tet promoter
Base Pairs: 56bp
Origin: Escherichia coli, synthetic
Properties: Inducible promoter regulated by tetracycline
Base Pairs: 56bp
Origin: Escherichia coli, synthetic
Properties: Inducible promoter regulated by tetracycline
== Usage and Biology ==
This is the naturally-occuring version of the TetR class B promoter. Note that this will promote
bidirectionally. (pR1 and pR2 will promote in the forward direction; pA will promote in the reverse
direction.) The three transcription start sites are not labeled, but can be found from the reference.
We set different tetracycline concentrations to test the protein expression induced by TetR(B) promoter. In our design, the immune protein (ike4) is regulated by TetR(B) promoter. The expression of the immune protein (ike4) directly affects and is positively correlated with strain growth. Therefore, the strain growth can indicate the regulation of the TetR(B) promoter.
The results of the test are below.
We set different tetracycline concentrations to test the protein expression induced by TetR(B) promoter. In our design, the immune protein (ike4) is regulated by TetR(B) promoter. The expression of the immune protein (ike4) directly affects and is positively correlated with strain growth. Therefore, the strain growth can indicate the regulation of the TetR(B) promoter.
The results of the test are below.
Table 1. OD600 value of the strain D: Pus232-ike4-tke4 after 6 hours
Figure 1. Biomass Concentration curve of Pus323-tke4-ike4 Strain with tetracycline
The results show that concentrations at the range of 300-500µg/L are more
effective. The results also proved that adding tetracycline to strain will boost the growth of strain. It
determines that the optimal concentration of tetracycline is 300µg/L to 500µg/L.
BBa_K4028006
== Profile ==
Name: tet pro-ike2
Base Pairs: 533 bp
Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic
Properties: Gene technology for protecting patented bacterial strains
Base Pairs: 533 bp
Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic
Properties: Gene technology for protecting patented bacterial strains
== Usage and Biology ==
A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs,
including opportunistic pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit
the human gut (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6]
Under natural conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or
antibacterial effects (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical
contact, thus inhibiting the growth of recipient cells. Meanwhile, the bacteria encoding T6SS can translate
and produce corresponding immune protein to counteract the damage caused by toxic effector factors.[1,2,3]
== Construct design ==
The immunity effector ike2 is under tet promoter. The composite part is inserted in
the pUS232 vector (Figure 3).
Figure 2. Immune protein expression box.
Figure 3. Schematic map of immune expression plasmids.
BBa_K4028007
== Profile ==
Name: tet pro-ike4
Base Pairs: 560 bp
Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic
Properties: Gene technology for protecting patented bacterial strains
Base Pairs: 560 bp
Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic
Properties: Gene technology for protecting patented bacterial strains
== Usage and Biology ==
A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs,
including opportunistic pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit
the human gut (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6]
Under natural conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or
antibacterial effects (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical
contact, thus inhibiting the growth of recipient cells. Meanwhile, the bacteria encoding T6SS can translate
and produce corresponding immune protein to counteract the damage caused by toxic effector factors.[1,2,3]
== Construct design ==
The immunity effector ike4 is under tet promoter(Figure 2). This composite part is
inserted in the pUS232 vector (Figure 3).
Figure 2. Immune protein expression box.
Figure 3. Schematic map of immune expression plasmids.
BBa_K4028008
== Profile ==
Name: lac pro-tke2-tet pro-ike2
Base Pairs: 4746 bp
Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic
Properties: Gene technology for protecting patented bacterial strains
Base Pairs: 4746 bp
Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic
Properties: Gene technology for protecting patented bacterial strains
== Usage and Biology ==
A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs,
including opportunistic pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit
the human gut (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6]
Under natural conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or
antibacterial effects (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical
contact, thus inhibiting the growth of recipient cells. Meanwhile, the bacteria encoding T6SS can translate
and produce corresponding immune protein to counteract the damage caused by toxic effector factors.[1,2,3]
== Construct design ==
With this idea coming from the T6SS, we designed to express the effector components
of T6SS in industrial strains by inserting the gene segment tke2-ike2 to induce the controlled expression of
the corresponding immune proteins. The normal growth of industrial microorganisms can be ensured only by
adding certain components (tetracycline) in the fermentation broth, so as to achieve the purpose of the
experiment.
Figure2. The gene design of TetR promoter-ike2-lac promoter-tke2.
Figure 3. Schematic map of T6SS effector and immune expression plasmids.
BBa_K4028009
== Profile ==
Name: lac pro-tke4-tet pro-ike4
Base Pairs: 5208 bp
Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic
Properties: Gene technology for protecting patented bacterial strains
Base Pairs: 5208 bp
Origin: Escherichia coli, Pseudomonas putida KT2440, synthetic
Properties: Gene technology for protecting patented bacterial strains
== Usage and Biology ==
A wide range of Gram-negative bacteria have been shown to have antibacterial T6SSs,
including opportunistic pathogens such as Pseudomonas aeruginosa,[4] obligate commensal species that inhabit
the human gut (Bacteroides spp.),[5] and plant-associated bacteria such as Agrobacterium tumefaciens.[6]
Under natural conditions, bacterial cells encoding T6SS transport effect factors with cytotoxic or
antibacterial effects (amidase, glycoside hydrolyase, lipase, etc.) to recipient cells through physical
contact, thus inhibiting the growth of recipient cells. Meanwhile, the bacteria encoding T6SS can translate
and produce corresponding immune protein to counteract the damage caused by toxic effector factors.[1,2,3]
== Construct design ==
Tke4 is a key functional factor that inhibits bacterial growth which is controlled
by a lac promoter (Figure 2). The immunity effector ike4 is under tet promoter. The anti-bacterial effector
tke4 and the immunity effector ike4 are inserted in the pUS232 vector (Figure 3).
Figure 2. T6SS effector and immune protein expression box.
Figure 3. Schematic map of T6SS effector and immune expression plasmids.