Part:BBa_K4028008
lac pro-tke2-tet pro-ike2
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 200
Illegal NgoMIV site found at 212
Illegal NgoMIV site found at 629
Illegal NgoMIV site found at 671
Illegal NgoMIV site found at 2920
Illegal AgeI site found at 2550 - 1000COMPATIBLE WITH RFC[1000]
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
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.
The profiles of every basic part are as follows:
BBa_K4028000
Name: tke2
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).
BBa_K4028001
Name: ike2
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). (T6SS).
BBa_K4028004
Name: lac promoter
Base Pairs: 55bp
Origin: Escherichia coli, synthetic
Properties: Constitutive promoter used for protein expression
Usage and Biology
This is a medium-strength Plac promoter. Placed before a gene, it promotes its expression. Plac is repressed by LacI. This promoter can be used for in vivo reporter systems; its medium strength is interesting for reporter genes that have a long half-life or that are not well supported by the cell.
BBa_K4028005
Name: tet promoter 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.
References
1. Bingle, l.E.H. et al. (2008). Type VI secretion: a beginner’s guide. Current opinion in microbiology. 11:3-8.
2. Silverman, J. M. et al. (2012). Structure and regulation of the type VI secretion system. 66:453-472.
3. Hernandez, R. E. et al. (2020). Type VI secretion system effector protein: Effective weapons for bacterial competitiveness. Cellular microbiology. 22:e13241.
4. Hood RD, . et al (January 2010). "A type VI secretion system ofPseudomonas aeruginosa targets a toxin to bacteria". Cell Host & Microbe. 7 (1): 25–37. doi:10.1016/j.chom.2009.12.007. PMC 2831478. PMID 20114026.
5. Russell AB, . et al (August 2014). "A type VI secretion-related pathway in Bacteroidetes mediates interbacterial antagonism". Cell Host & Microbe. 16 (2): 227–236. doi:10.1016/j.chom.2014.07.007. PMC 4136423. PMID 25070807.
6. Ma LS, . et al (July 2014). "Agrobacterium tumefaciens deploys a superfamily of type VI secretion DNase effectors as weapons for interbacterial competition in planta". Cell Host & Microbe. 16 (1): 94–104. doi:10.1016/j.chom.2014.06.002. PMC 4096383. PMID 24981331.
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