Difference between revisions of "Part:BBa K5267011"

 
 
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<partinfo>BBa_K5267011 short</partinfo>
 
<partinfo>BBa_K5267011 short</partinfo>
  
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A signal peptide is a hydrophobic amino acid sequence located at the N-terminus of a protein encoding a length of 5-30 amino acids used to direct newly synthesised proteins towards the endoplasmic reticulum. Signal peptides are found in secreted proteins, transmembrane proteins, and proteins within eukaryotic organelles.
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promoter that responsible for transcription of MTNR1a
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
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<partinfo>BBa_K5267011 parameters</partinfo>
 
<partinfo>BBa_K5267011 parameters</partinfo>
 
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===Profile===
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Name: mPGK promoter
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<br>Base Pairs: 516bp
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<br>Origin: Mouse
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<br>Properties: promoter that responsible for transcription of MTNR1a
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===Usage and Biology===
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MPGK promoter(mpPGK) is the promoter of Pgk1 in mouse cell. Pgk-1 is the murine gene encoding phosphoglycerate kinase (PGK), a key enzyme in glycolysis. This enzyme must  be present in all cells, so the Pgk-1 promoter is expected to be a ubiquitously active one<sup>[1]</sup>.
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The cloned mpPGK is very active in driving transcription of reporter genes following transfection into mammalian cells<sup>[2]</sup>. The promoter is particularly active in pluripotent embryonal carcinoma and embryonic stem cells.
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<br>An initial studie on the Pgk-1 promoter indicated that its activity was contained within a region from −425 bp to +80 bp relative to the first transcription start site[2]. This promoter is GC rich and contains no TATA box<sup>[3]</sup>. The data from John P. Thomson et al. indicate that a primary function of non-methylated CGIs is to genetically influence the local chromatin modification state to initiate transcriptionby interaction with Cfp1 and perhaps other CpG-binding proteins<sup>[4]</sup>. The core promoter from −121 bp to +80 bp contains a number of putative Spl binding sites and had modest promoter activity. The 304 bp upstream of the core promoter had enhancer-like activity such that this region increased expression from the core promoter in an orientation- and position-independent fashion.
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===Figure===
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<html>
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<figure class="figure">
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<div style="width=100%;height=auto;align-items:center">
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<img src="https://static.igem.wiki/teams/5267/i-m-zhangrenjie/1.jpg" class="figure-img img-fluid rounded"  height="200px">
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<img src="https://static.igem.wiki/teams/5267/i-m-zhangrenjie/2.jpg" class="figure-img img-fluid rounded"  height="200px">
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</figure>
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</html>
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<br>Regulated expression from the Pgk-1 promoter carrying the UAS. The Pgk-1 promoter comprising the region spanning −425 to +80 bp (black bars) and −121 to +80 bp were used to drive the lacZ reporter gene. (A) These constructs were cotransfected into P19 cells along with an RSV-driven CAT construct and harvested 48 h later. The P19 cells were treated with retinoic acid (RA) for the number of days shown before cells were harvested for measurement of β-galactosidase and CAT activities. (B) The two test constructs were cotransfected along with a selectible gene, Pgk-puro, and stably transformed cells were selected in puromycin. Colonies of transformed cells were pooled and treated with RA for the number of days shown before being harvested for β-galactosidase and CAT activities. The activity shown is that of β-galactosidase normalized to the CAT control.[1]
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===Sequence===
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Top:
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<br>GGAGCTACCGGGTAGGGGAGGCGCTTTTCCCAAGGCAGTCTGGAGCATGCGCTTTAGCAGCCCCGCTGGGCACTTGGCGCTACACAAGTGGC
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<br>CTCTGGCCTCGCACACATTCCACATCCACCGGTAGCGCCAACCGGCTCCGTTCTTTGGTGGCCCCTTCGCGCCACCTTCTACTCCTCCCCTA
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<br>GTCAGGAAGTTCCCCCCCGCCCCGCAGCTCGCGTCGTGCAGGACGTGACAAATGGAAGTAGCACGTCTCAGTAGTCTCGTGCAGATGGACAG
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<br>CACCGCTGAGCAATGGAAGCGGGTAGGCCTTTGGGGCAGCGGCCAATAGCAGCTTTGCTCCTTCGCTTTCTGGGCTCAGAGGCTGGGAAGGG
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<br>GTGGGTCCGGGGGCGGGCTCAGGGGCGGGCTCAGGGGCGGGGCGGGCGCCCGAAGGTCCTCCGGAGGCCCGGCATTCTGCACGCTTCAAAAG
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<br>CGCACGTCTGCCGCGCTGTTCTCCTCTTCCTCATCTCCGGGCCTTTCGACCTCCAGCCCTACT
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===Reference===
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[1] SUTHERLAND L C, ST-ARNAUD R, MCBURNEY M W. An upstream activator sequence regulates the murine Pgk-1 promoter and binds multiple nuclear proteins [J]. Gene expression, 1995, 4(4-5): 265-79.
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<br>[2] MCBURNEY M W, SUTHERLAND L C, ADRA C N, et al. The mouse Pgk-1 gene promoter contains an upstream activator sequence [J]. Nucleic acids research, 1991, 19(20): 5755-61.
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<br>[3] ADRA C N, BOER P H, MCBURNEY M W. Cloning and expression of the mouse pgk-1 gene and the nucleotide sequence of its promoter [J]. Gene, 1987, 60(1): 65-74.
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<br>[4] THOMSON J P, SKENE P J, SELFRIDGE J, et al. CpG islands influence chromatin structure via the CpG-binding protein Cfp1 [J]. Nature, 2010, 464(7291): 1082-U162.

Latest revision as of 17:58, 25 September 2024


mPGK Promoter

promoter that responsible for transcription of MTNR1a Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 116
  • 1000
    COMPATIBLE WITH RFC[1000]


Profile

Name: mPGK promoter
Base Pairs: 516bp
Origin: Mouse
Properties: promoter that responsible for transcription of MTNR1a


Usage and Biology

MPGK promoter(mpPGK) is the promoter of Pgk1 in mouse cell. Pgk-1 is the murine gene encoding phosphoglycerate kinase (PGK), a key enzyme in glycolysis. This enzyme must be present in all cells, so the Pgk-1 promoter is expected to be a ubiquitously active one[1]. The cloned mpPGK is very active in driving transcription of reporter genes following transfection into mammalian cells[2]. The promoter is particularly active in pluripotent embryonal carcinoma and embryonic stem cells.
An initial studie on the Pgk-1 promoter indicated that its activity was contained within a region from −425 bp to +80 bp relative to the first transcription start site[2]. This promoter is GC rich and contains no TATA box[3]. The data from John P. Thomson et al. indicate that a primary function of non-methylated CGIs is to genetically influence the local chromatin modification state to initiate transcriptionby interaction with Cfp1 and perhaps other CpG-binding proteins[4]. The core promoter from −121 bp to +80 bp contains a number of putative Spl binding sites and had modest promoter activity. The 304 bp upstream of the core promoter had enhancer-like activity such that this region increased expression from the core promoter in an orientation- and position-independent fashion.


Figure


Regulated expression from the Pgk-1 promoter carrying the UAS. The Pgk-1 promoter comprising the region spanning −425 to +80 bp (black bars) and −121 to +80 bp were used to drive the lacZ reporter gene. (A) These constructs were cotransfected into P19 cells along with an RSV-driven CAT construct and harvested 48 h later. The P19 cells were treated with retinoic acid (RA) for the number of days shown before cells were harvested for measurement of β-galactosidase and CAT activities. (B) The two test constructs were cotransfected along with a selectible gene, Pgk-puro, and stably transformed cells were selected in puromycin. Colonies of transformed cells were pooled and treated with RA for the number of days shown before being harvested for β-galactosidase and CAT activities. The activity shown is that of β-galactosidase normalized to the CAT control.[1]


Sequence

Top:
GGAGCTACCGGGTAGGGGAGGCGCTTTTCCCAAGGCAGTCTGGAGCATGCGCTTTAGCAGCCCCGCTGGGCACTTGGCGCTACACAAGTGGC
CTCTGGCCTCGCACACATTCCACATCCACCGGTAGCGCCAACCGGCTCCGTTCTTTGGTGGCCCCTTCGCGCCACCTTCTACTCCTCCCCTA
GTCAGGAAGTTCCCCCCCGCCCCGCAGCTCGCGTCGTGCAGGACGTGACAAATGGAAGTAGCACGTCTCAGTAGTCTCGTGCAGATGGACAG
CACCGCTGAGCAATGGAAGCGGGTAGGCCTTTGGGGCAGCGGCCAATAGCAGCTTTGCTCCTTCGCTTTCTGGGCTCAGAGGCTGGGAAGGG
GTGGGTCCGGGGGCGGGCTCAGGGGCGGGCTCAGGGGCGGGGCGGGCGCCCGAAGGTCCTCCGGAGGCCCGGCATTCTGCACGCTTCAAAAG
CGCACGTCTGCCGCGCTGTTCTCCTCTTCCTCATCTCCGGGCCTTTCGACCTCCAGCCCTACT

Reference

[1] SUTHERLAND L C, ST-ARNAUD R, MCBURNEY M W. An upstream activator sequence regulates the murine Pgk-1 promoter and binds multiple nuclear proteins [J]. Gene expression, 1995, 4(4-5): 265-79.
[2] MCBURNEY M W, SUTHERLAND L C, ADRA C N, et al. The mouse Pgk-1 gene promoter contains an upstream activator sequence [J]. Nucleic acids research, 1991, 19(20): 5755-61.
[3] ADRA C N, BOER P H, MCBURNEY M W. Cloning and expression of the mouse pgk-1 gene and the nucleotide sequence of its promoter [J]. Gene, 1987, 60(1): 65-74.
[4] THOMSON J P, SKENE P J, SELFRIDGE J, et al. CpG islands influence chromatin structure via the CpG-binding protein Cfp1 [J]. Nature, 2010, 464(7291): 1082-U162.