Difference between revisions of "Part:BBa K3970025"

 
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<partinfo>BBa_K3970025 short</partinfo>
 
<partinfo>BBa_K3970025 short</partinfo>
  
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Gene PcyA encodes ferredoxin oxidoreductase, which catalyzes the four-electron reduction of biliverdin IX-alpha (2-electron reduction at both the A and D rings) and then form (2R,3Z)-phycocyanobilin:
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==Usage and Biology==
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biliverdin IX-alpha + 4 H+ + 4 reduced [2Fe-2S]-[ferredoxin] = biliverdin IX-alpha + 4 oxidized [2Fe-2S]-[ferredoxin]
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CpcA gene encodes α-phycocyanin, a light-harvesting photosynthetic bile pigment-protein from the phycobiliprotein complex (phycobilisome, PBS). Phycocyanin is the major phycobiliprotein in the PBS rod.
  
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<!-- Add more about the biology of this part here
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==Construction==
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===Usage and Biology===
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We use primer:XbaI-GAP-up and primer:GAP-cpcA-dn to get the fragment of GAP.
  
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We use primer:cpcA-up and primer:cpcA-ADH1-dn to get the fragment of cpcA.
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We use primer:ADH1-up and primer:ADH1-XbaI-dn to get the fragment of ADH1.
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[[File:0024-1.png|600px|thumb|center]]
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[[File:0024-2.png|600px|thumb|center]]
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<b>Primers we use to construct cpcA expression cassette:</b>
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XbaI-GAP-up (48-mer): tcgctccccatttctctagtcattatcaatactgccatttcaaagaat
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GAP-cpcA-dn (40-mer): ttcggtgataggggtcttcatggtggcgagatctaattcg
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cpcA-up (23-mer): atgaagacccctatcaccgaagc
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cpcA-ADH1-dn (41-mer): tcataaatcataagaaattcgctcaggatagagcattgata
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ADH1-up (30-mer): gcgaatttcttatgatttatgatttttatt
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ADH1-XbaI-dn (33-mer): Cgaactctgcagtcttatatgccggtagaggt
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==Experiment results==
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the experimental pictures shows how we recycle the fragment of ADH1 terminator.
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( note that how we recycle the GAP promoter and the gene cpcaA has been shown on the last parts.)
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[[File:0024-3.png|400px|thumb|left|Fig. The fragments of the constructed plasmid were recovered]]
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<br><br><br>
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<b>M:  Marker </b>
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<b>1-4: ADH1(terminator)</b>
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<br><br><br><br><br><br><br><br><br><br>
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>

Revision as of 05:14, 7 October 2021


pGAP-PcyA-tADH1

Usage and Biology

CpcA gene encodes α-phycocyanin, a light-harvesting photosynthetic bile pigment-protein from the phycobiliprotein complex (phycobilisome, PBS). Phycocyanin is the major phycobiliprotein in the PBS rod.

Construction

We use primer:XbaI-GAP-up and primer:GAP-cpcA-dn to get the fragment of GAP.

We use primer:cpcA-up and primer:cpcA-ADH1-dn to get the fragment of cpcA.

We use primer:ADH1-up and primer:ADH1-XbaI-dn to get the fragment of ADH1.

0024-1.png
0024-2.png

Primers we use to construct cpcA expression cassette:

XbaI-GAP-up (48-mer): tcgctccccatttctctagtcattatcaatactgccatttcaaagaat

GAP-cpcA-dn (40-mer): ttcggtgataggggtcttcatggtggcgagatctaattcg

cpcA-up (23-mer): atgaagacccctatcaccgaagc

cpcA-ADH1-dn (41-mer): tcataaatcataagaaattcgctcaggatagagcattgata

ADH1-up (30-mer): gcgaatttcttatgatttatgatttttatt

ADH1-XbaI-dn (33-mer): Cgaactctgcagtcttatatgccggtagaggt

Experiment results

the experimental pictures shows how we recycle the fragment of ADH1 terminator. ( note that how we recycle the GAP promoter and the gene cpcaA has been shown on the last parts.)

Fig. The fragments of the constructed plasmid were recovered







M: Marker

1-4: ADH1(terminator)









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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]