Difference between revisions of "Part:BBa K3993013"
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<partinfo>BBa_K3993013 short</partinfo> | <partinfo>BBa_K3993013 short</partinfo> | ||
| − | PPGK1-AsADC-TADH1 - PTEF1-SPE1-TTEF1 - PTDH3-speB-TCYC1 | + | === Profile === |
| + | ====Name: PPGK1-AsADC-TADH1 - PTEF1-SPE1-TTEF1 - PTDH3-speB-TCYC1==== | ||
| + | ====Base Pairs: 6866 bp==== | ||
| + | ====Origin: Saccharomyces cerevisiae, E. coli, synthesis, genome==== | ||
| + | ====Properties: Arginine metabolism and polyamine biosynthesis chemical reactions==== | ||
| + | |||
| + | === Usage and Biology === | ||
| + | Kl tropane alkaloids (TAs) refers to a kind of alkaloids containing the tropane alkyl skeleton formed by the combination of pyrrole ring and piperidine ring in structure. It is a natural product of plant and has a long history and important medicinal value. tropane alkaloids have great market demand and often appear in global shortages. A method that can produce Tas in scale is expected. Using synthetic biology to create a microbial cell factory to produce TAs is a highly potential strategy. | ||
| + | |||
| + | The Tropane alkaloid (TAs) is obtained by a series of chemical reactions through the formation of Putrescine (1, 4-butylenediamine, Putrescine) from Arginine. Putrescine is an essential polyamine for ribosomal biogenesis and mRNA translation, but is regulated by polyamines and remains at low concentrations during normal cell growth. In this study, by overexpressing the natural genes involved in arginine metabolism and polyamine biosynthesis, the regulatory mechanism of polyamine biosynthesis is adjusted, so as to engineer the production of excessive putrescine strains. | ||
| + | |||
| + | [[File:T--SHSID--BBa K3993000 Figure1.png|500px|thumb|center|Figure1. Principle diagram of TAs..]] | ||
| + | |||
| + | === Construct design === | ||
| + | The Tropine part of Tropane alkaloids (TAs) is obtained from arginine to putrescine (1,4-butanediamine, putrescine), and then through a series of chemical reactions. In this project, natural genes involved in arginine metabolism and polyamine biosynthesis was designed to overexpress in yeast. The engineer strains that produced excess putrescine. (Figure 2). | ||
| + | |||
| + | [[File:T--SHSID--BBa K3993013-Figure2.png|500px|thumb|center|Figure 2. DNA sequence map of plasmid pYES2-AsADC-SPE1-SpeB..]] | ||
| + | |||
| + | ===The profiles of every basic part are as follows:=== | ||
| + | == BBa_K3993000== | ||
| + | ====Name: SPE1==== | ||
| + | ====Base Pairs: 1401bp==== | ||
| + | ====Origin: Saccharomyces cerevisiae, genome==== | ||
| + | ====Properties: Catalyzes the first and rate-limiting step of polyamine biosynthesis that converts ornithine into putrescine==== | ||
| + | |||
| + | === Usage and Biology === | ||
| + | his protein is involved in step 1 of the subpathway that synthesizes putrescine from L-ornithine. Catalyzes the first and rate-limiting step of polyamine biosynthesis that converts ornithine into putrescine, which is the precursor for the polyamines, spermidine and spermine. Polyamines are essential for cell proliferation and are implicated in cellular processes, ranging from DNA replication to apoptosis. Homodimer and only the dimer is catalytically active, as the active sites are constructed of residues from both monomers | ||
| + | |||
| + | |||
| + | == BBa_K3993001 == | ||
| + | ====Name: SPEB==== | ||
| + | ====Base Pairs: 921bp==== | ||
| + | ====Origin: E. coli, genome==== | ||
| + | ====Properties: Catalyzes the formation of putrescine from agmatine.==== | ||
| + | |||
| + | === Usage and Biology === | ||
| + | This protein is involved in step 1 of the subpathway that synthesizes putrescine from agmatine. This subpathway is part of the pathway putrescine biosynthesis via agmatine pathway, which is itself part of Amine and polyamine biosynthesis. The expression of AUH activity is antagonistically regulated by cyclic AMP and agmatine. In the presence of the cAMP receptor protein, cAMP represses the expression of AUH, while agmatine induces it. | ||
| + | |||
| + | |||
| + | == BBa_K3993002== | ||
| + | ====Name: AsADC==== | ||
| + | ====Base Pairs: 1821bp==== | ||
| + | ====Origin: Saccharomyces cerevisiae, synthesis==== | ||
| + | ====Properties: codon optimized SPE1==== | ||
| + | |||
| + | === Usage and Biology === | ||
| + | This protein is involved in step 1 of the subpathway that synthesizes agmatine from L-arginine. This subpathway is part of the pathway agmatine biosynthesis, which is itself part of Amine and polyamine biosynthesis. | ||
| + | |||
| + | == BBa_K3993003 == | ||
| + | ====Name: PTDH3==== | ||
| + | ====Base Pairs: 673bp==== | ||
| + | ====Origin: Addgene==== | ||
| + | ====Properties: Yeast centromeric vector with the TDH3 (glyceraldehyde 3-phosphate dehydrogenase) promoter.==== | ||
| + | |||
| + | === Usage and Biology === | ||
| + | Yeast CEN/ARS vector (Leu2) that contains multiple cloning site ( MCS ) and TDH3 promoter. | ||
| + | |||
| + | == BBa_K3993004 == | ||
| + | ====Name: TCYC1==== | ||
| + | ====Base Pairs: 242bp==== | ||
| + | ====Origin: Saccharomyces cerevisiae, genome==== | ||
| + | ====Properties: CYC1 terminator==== | ||
| + | |||
| + | === Usage and Biology === | ||
| + | This is a common transcriptional terminator. Placed after a gene, it completing the transcription process and impacting mRNA half-life. This terminator can be used for in vivo systems, and can be used for modulating gene expression in yeast. | ||
| + | |||
| + | == BBa_K3993005 == | ||
| + | ====Name: PPGK1==== | ||
| + | ====Base Pairs: 502bp==== | ||
| + | ====Origin: Addgene==== | ||
| + | ====Properties: A Yeast Expression plasmids backbone==== | ||
| + | |||
| + | === Usage and Biology === | ||
| + | Selection for in-frame fusion expression constructs. | ||
| + | |||
| + | |||
| + | == BBa_K3993006== | ||
| + | ====Name: TADH1==== | ||
| + | ====Base Pairs: 328bp==== | ||
| + | ====Origin: Saccharomyces cerevisiae, genome==== | ||
| + | ====Properties: ADH1 terminator==== | ||
| + | |||
| + | === Usage and Biology === | ||
| + | This is a common transcriptional terminator. Placed after a gene, it completing the transcription process and impacting mRNA half-life. This terminator can be used for in vivo systems, and can be used for modulating gene expression in yeast. | ||
| + | |||
| + | |||
| + | |||
| + | |||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Revision as of 02:24, 20 October 2021
PPGK1-AsADC-TADH1 - PTEF1-SPE1-TTEF1 - PTDH3-speB-TCYC1
Profile
Name: PPGK1-AsADC-TADH1 - PTEF1-SPE1-TTEF1 - PTDH3-speB-TCYC1
Base Pairs: 6866 bp
Origin: Saccharomyces cerevisiae, E. coli, synthesis, genome
Properties: Arginine metabolism and polyamine biosynthesis chemical reactions
Usage and Biology
Kl tropane alkaloids (TAs) refers to a kind of alkaloids containing the tropane alkyl skeleton formed by the combination of pyrrole ring and piperidine ring in structure. It is a natural product of plant and has a long history and important medicinal value. tropane alkaloids have great market demand and often appear in global shortages. A method that can produce Tas in scale is expected. Using synthetic biology to create a microbial cell factory to produce TAs is a highly potential strategy.
The Tropane alkaloid (TAs) is obtained by a series of chemical reactions through the formation of Putrescine (1, 4-butylenediamine, Putrescine) from Arginine. Putrescine is an essential polyamine for ribosomal biogenesis and mRNA translation, but is regulated by polyamines and remains at low concentrations during normal cell growth. In this study, by overexpressing the natural genes involved in arginine metabolism and polyamine biosynthesis, the regulatory mechanism of polyamine biosynthesis is adjusted, so as to engineer the production of excessive putrescine strains.
Construct design
The Tropine part of Tropane alkaloids (TAs) is obtained from arginine to putrescine (1,4-butanediamine, putrescine), and then through a series of chemical reactions. In this project, natural genes involved in arginine metabolism and polyamine biosynthesis was designed to overexpress in yeast. The engineer strains that produced excess putrescine. (Figure 2).
The profiles of every basic part are as follows:
BBa_K3993000
Name: SPE1
Base Pairs: 1401bp
Origin: Saccharomyces cerevisiae, genome
Properties: Catalyzes the first and rate-limiting step of polyamine biosynthesis that converts ornithine into putrescine
Usage and Biology
his protein is involved in step 1 of the subpathway that synthesizes putrescine from L-ornithine. Catalyzes the first and rate-limiting step of polyamine biosynthesis that converts ornithine into putrescine, which is the precursor for the polyamines, spermidine and spermine. Polyamines are essential for cell proliferation and are implicated in cellular processes, ranging from DNA replication to apoptosis. Homodimer and only the dimer is catalytically active, as the active sites are constructed of residues from both monomers
BBa_K3993001
Name: SPEB
Base Pairs: 921bp
Origin: E. coli, genome
Properties: Catalyzes the formation of putrescine from agmatine.
Usage and Biology
This protein is involved in step 1 of the subpathway that synthesizes putrescine from agmatine. This subpathway is part of the pathway putrescine biosynthesis via agmatine pathway, which is itself part of Amine and polyamine biosynthesis. The expression of AUH activity is antagonistically regulated by cyclic AMP and agmatine. In the presence of the cAMP receptor protein, cAMP represses the expression of AUH, while agmatine induces it.
BBa_K3993002
Name: AsADC
Base Pairs: 1821bp
Origin: Saccharomyces cerevisiae, synthesis
Properties: codon optimized SPE1
Usage and Biology
This protein is involved in step 1 of the subpathway that synthesizes agmatine from L-arginine. This subpathway is part of the pathway agmatine biosynthesis, which is itself part of Amine and polyamine biosynthesis.
BBa_K3993003
Name: PTDH3
Base Pairs: 673bp
Origin: Addgene
Properties: Yeast centromeric vector with the TDH3 (glyceraldehyde 3-phosphate dehydrogenase) promoter.
Usage and Biology
Yeast CEN/ARS vector (Leu2) that contains multiple cloning site ( MCS ) and TDH3 promoter.
BBa_K3993004
Name: TCYC1
Base Pairs: 242bp
Origin: Saccharomyces cerevisiae, genome
Properties: CYC1 terminator
Usage and Biology
This is a common transcriptional terminator. Placed after a gene, it completing the transcription process and impacting mRNA half-life. This terminator can be used for in vivo systems, and can be used for modulating gene expression in yeast.
BBa_K3993005
Name: PPGK1
Base Pairs: 502bp
Origin: Addgene
Properties: A Yeast Expression plasmids backbone
Usage and Biology
Selection for in-frame fusion expression constructs.
BBa_K3993006
Name: TADH1
Base Pairs: 328bp
Origin: Saccharomyces cerevisiae, genome
Properties: ADH1 terminator
Usage and Biology
This is a common transcriptional terminator. Placed after a gene, it completing the transcription process and impacting mRNA half-life. This terminator can be used for in vivo systems, and can be used for modulating gene expression in yeast.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 6016
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 2321
Illegal PstI site found at 6016 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1191
Illegal BglII site found at 1412
Illegal BglII site found at 4342
Illegal BglII site found at 6492
Illegal BamHI site found at 6405 - 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 6016
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 6016
Illegal AgeI site found at 6427 - 1000COMPATIBLE WITH RFC[1000]