SquarePants (Talk | contribs) |
SquarePants (Talk | contribs) |
||
Line 30: | Line 30: | ||
<h2 style="margin:30px; color:#FFF"><br> Parts Overview</h2> | <h2 style="margin:30px; color:#FFF"><br> Parts Overview</h2> | ||
<p style="margin:30px; color:#FFF"> We have registered 31 basic parts (BBa_ K3100001-BBa_K310003) and 41 composite parts (BBa_K3100100-BBa_K3100140), all of which is BioBrick RFC10 or Type IIS compatible.<br> | <p style="margin:30px; color:#FFF"> We have registered 31 basic parts (BBa_ K3100001-BBa_K310003) and 41 composite parts (BBa_K3100100-BBa_K3100140), all of which is BioBrick RFC10 or Type IIS compatible.<br> | ||
− | BBa_K3100001 to BBa_K3100010 is different strength T7 promoter which is from the research of Amar Ghodasara<em>et al</em> <sup>[1]</sup>. Toehold Switch is prokaryotic riboregulators<sup>[2]</sup>, we have added Toehold Switch A&C and their Trigger DNAs (Toehold Switch B&D is from previous part number range). BBa_K3100100-BBa_K3100139 are composite parts which are the combination of 4 different Triggers and 10 unique T7 promoters. Through specific design of Fusion Sites, these 40 parts will be randomly combined by Golden Gate assembly. Up to 4 different Triggers driven by 10 unique promoters are expressed from one plasmid and finally a pool called VerProS with a storage capacity of 10000 is obtained. VerProS pool can simultaneously optimize up to four genes in a system. Particularly, this versatile library can be applied to fast optimization in different systems without having to build <em>ad hoc</em> libraries, which can greatly reduce manpower and costs.<br> | + | BBa_K3100001 to BBa_K3100010 is different strength T7 promoter which is from the research of Amar Ghodasara<em>et al</em> <sup>[1]</sup>. Toehold Switch is prokaryotic riboregulators<sup>[2]</sup>, we have added Toehold Switch A&C and their Trigger DNAs (Toehold Switch B&D is from previous part number range). BBa_K3100100-BBa_K3100139 are composite parts which are the combination of 4 different Triggers and 10 unique T7 promoters. Through specific design of Fusion Sites, these 40 parts will be randomly combined by Golden Gate assembly. Up to 4 different Triggers driven by 10 unique promoters are expressed from one plasmid and finally a pool called VerProS with a storage capacity of 10000 is obtained. VerProS pool can simultaneously optimize up to four genes in a system. Particularly, this versatile library can be applied to fast optimization in different systems without having to build <em>ad hoc</em> libraries, which can greatly reduce manpower and costs.<br> |
We have added 4 acid tolerant factors (BBa_K3100017- BBa_K3100020) and have tested their influence on the acid tolerance of <em>E.coli</em>. What’s more, BBa_K3100140 is a composite part combined of 4 acid tolerant factor and 4 Toehold Switch. In our Project, we use the VerProS pool for fine regulation of these 4 acid tolerant factors.<br> | We have added 4 acid tolerant factors (BBa_K3100017- BBa_K3100020) and have tested their influence on the acid tolerance of <em>E.coli</em>. What’s more, BBa_K3100140 is a composite part combined of 4 acid tolerant factor and 4 Toehold Switch. In our Project, we use the VerProS pool for fine regulation of these 4 acid tolerant factors.<br> | ||
As T7 is the most well-known inducible promoter with high transcriptional strength <sup>[3]</sup>, we want to construct and improve a T7 promoter library which contains more T7 promoters with different strength. Therefore, the library can help our VerProS system to achieve more fine-grained regulation. Site-saturation mutagenesis was conducted to obtain the T7 mutants.BBa_K3100022-BBa_K3100031 is the different strength T7 promoters which we have constructed and improved. <br><br>[1] Ghodasara A, Voigt CA. (2017) Balancing gene expression without library construction via a reusable sRNA pool.<br>[2]Green AA, Silver PA, Collins JJ, Yin P. (2014) Toehold switches: <em>de-novo</em>-designed regulators of gene expression.<br>[3] Nie, Z., Luo, H., Li, J. et al. Appl Biochem Biotechnol (2019). https://doi.org/10.1007/s12010-019-03113-y</p><br></div> | As T7 is the most well-known inducible promoter with high transcriptional strength <sup>[3]</sup>, we want to construct and improve a T7 promoter library which contains more T7 promoters with different strength. Therefore, the library can help our VerProS system to achieve more fine-grained regulation. Site-saturation mutagenesis was conducted to obtain the T7 mutants.BBa_K3100022-BBa_K3100031 is the different strength T7 promoters which we have constructed and improved. <br><br>[1] Ghodasara A, Voigt CA. (2017) Balancing gene expression without library construction via a reusable sRNA pool.<br>[2]Green AA, Silver PA, Collins JJ, Yin P. (2014) Toehold switches: <em>de-novo</em>-designed regulators of gene expression.<br>[3] Nie, Z., Luo, H., Li, J. et al. Appl Biochem Biotechnol (2019). https://doi.org/10.1007/s12010-019-03113-y</p><br></div> |
Revision as of 13:11, 18 October 2019
Parts Overview
We have registered 31 basic parts (BBa_ K3100001-BBa_K310003) and 41 composite parts (BBa_K3100100-BBa_K3100140), all of which is BioBrick RFC10 or Type IIS compatible.
BBa_K3100001 to BBa_K3100010 is different strength T7 promoter which is from the research of Amar Ghodasaraet al [1]. Toehold Switch is prokaryotic riboregulators[2], we have added Toehold Switch A&C and their Trigger DNAs (Toehold Switch B&D is from previous part number range). BBa_K3100100-BBa_K3100139 are composite parts which are the combination of 4 different Triggers and 10 unique T7 promoters. Through specific design of Fusion Sites, these 40 parts will be randomly combined by Golden Gate assembly. Up to 4 different Triggers driven by 10 unique promoters are expressed from one plasmid and finally a pool called VerProS with a storage capacity of 10000 is obtained. VerProS pool can simultaneously optimize up to four genes in a system. Particularly, this versatile library can be applied to fast optimization in different systems without having to build ad hoc libraries, which can greatly reduce manpower and costs.
We have added 4 acid tolerant factors (BBa_K3100017- BBa_K3100020) and have tested their influence on the acid tolerance of E.coli. What’s more, BBa_K3100140 is a composite part combined of 4 acid tolerant factor and 4 Toehold Switch. In our Project, we use the VerProS pool for fine regulation of these 4 acid tolerant factors.
As T7 is the most well-known inducible promoter with high transcriptional strength [3], we want to construct and improve a T7 promoter library which contains more T7 promoters with different strength. Therefore, the library can help our VerProS system to achieve more fine-grained regulation. Site-saturation mutagenesis was conducted to obtain the T7 mutants.BBa_K3100022-BBa_K3100031 is the different strength T7 promoters which we have constructed and improved.
[1] Ghodasara A, Voigt CA. (2017) Balancing gene expression without library construction via a reusable sRNA pool.
[2]Green AA, Silver PA, Collins JJ, Yin P. (2014) Toehold switches: de-novo-designed regulators of gene expression.
[3] Nie, Z., Luo, H., Li, J. et al. Appl Biochem Biotechnol (2019). https://doi.org/10.1007/s12010-019-03113-y
Parts Table
(这里放表格)