Difference between revisions of "Part:BBa K2374003"
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It belongs to the biopterin-dependent aromatic amino acid hydroxylase family. We also provide ple promoter in [https://parts.igem.org/Part:BBa_K2374001 BBa_K2374001]. | It belongs to the biopterin-dependent aromatic amino acid hydroxylase family. We also provide ple promoter in [https://parts.igem.org/Part:BBa_K2374001 BBa_K2374001]. | ||
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+ | We construct pSB1C3-UAS-TH and pUAST-UAS-TH. The pSB1C3-UAS-TH is for submission. Then pUAST-UAS-TH is used to do micro-injection into the D.melanogaster. Also with the other two plasmids: pUAST-ple-GAL4 ([https://parts.igem.org/Part:BBa_K2374005 BBa_K2374005])and pUAST-ple-GAL80ts ([https://parts.igem.org/Part:BBa_K2374006 BBa_K2374006]). We must combine the three pathways to determine if the system work well. The result of our testing on D.melanogaster(pUAST-UAS-TH) is displayed below. | ||
Revision as of 18:51, 29 October 2017
ple (Tyrosine 3-monooxygenase, TH) -> (fruit fly)
UAS-TH | |
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Use in | D.melanogaster |
RFC standard | RFC 10 compatible |
Backbone | pSB1C3 |
Submitted by | [http://2017.igem.org/Team:Tongji_China Tongji_China 2017] |
An upstream activating sequence or upstream activation sequence (UAS) is a cis-acting regulatory sequence. It is distinct from the promoter and increases the expression of a neighbouring gene. Due to its essential role in activating transcription, the UAS is often considered to be analogous to the function of the enhancer in multicellular eukaryotes. Upstream activation sequences are a crucial part of induction, enhancing the expression of the protein of interest through increased transcriptional activity. The UAS is found adjacently upstream to a minimal promoter (TATA box) and serves as a binding site for transactivators. If the transcriptional transactivator does not bind to the UAS in the proper orientation then transcription cannot begin
Gene ple encodes Tyrosine 3-monooxygenase which also known as TH. It plays an important role in the physiology of adrenergic neurons. This protein is involved in step 1 of the subpathway that synthesizes dopamine from L-tyrosine. Dopamine has critical roles in system development. Proteins known to be involved in the 2 steps of the subpathway in this organism are: 1.Tyrosine 3-monooxygenase (ple) 2.no protein annotated in this organism This subpathway is part of the pathway dopamine biosynthesis, which is itself part of Catecholamine biosynthesis. It belongs to the biopterin-dependent aromatic amino acid hydroxylase family. We also provide ple promoter in BBa_K2374001.
We construct pSB1C3-UAS-TH and pUAST-UAS-TH. The pSB1C3-UAS-TH is for submission. Then pUAST-UAS-TH is used to do micro-injection into the D.melanogaster. Also with the other two plasmids: pUAST-ple-GAL4 (BBa_K2374005)and pUAST-ple-GAL80ts (BBa_K2374006). We must combine the three pathways to determine if the system work well. The result of our testing on D.melanogaster(pUAST-UAS-TH) is displayed below.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1157
Illegal XhoI site found at 289 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 535
Illegal BsaI site found at 1501
Illegal BsaI.rc site found at 253
Illegal BsaI.rc site found at 1019
Illegal BsaI.rc site found at 1168
Functional Parameters
TH is involved in step 1 of the subpathway that synthesizes dopamine from L-tyrosine and plays an important role in the physiology of adrenergic neurons.
References
1. Webster, Nocholas; Jin, Jia Rui; Green, Stephen; Hollis, Melvyn; Chambon, Pierre (29 January 1988). "The Yeast UASG is a transcriptional enhancer in human hela cells in the presence of the GAL4 trans-activator". Cell. 52 (2): 169–178.
2. West, Jr., Robert W.; Yocum, R. Rogers; Ptashne, Mark (November 1984). "Saccharomyces cerevisiae GAL1-GAL10 Divergenet Promoter Region: Location and Function of the Upstream Activating Sequence UASG". Molecular and Cellular Biology. 4 (11): 2467–2478.
3. Lewandoski, Mark (October 2001). "Conditional control of gene expression in the mouse". Nature Reviews Genetics. 2: 743–755.