Difference between revisions of "Part:BBa K3970024"
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<partinfo>BBa_K3970024 short</partinfo> | <partinfo>BBa_K3970024 short</partinfo> | ||
− | CpcA gene encodes | + | ==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. | ||
+ | |||
+ | [[File:0024-1.png|600px|thumb|center]] | ||
+ | |||
+ | [[File:0024-2.png|600px|thumb|center]] | ||
+ | |||
+ | <b>Primers we use to construct cpcA expression cassette:</b> | ||
+ | |||
+ | 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.) | ||
+ | |||
+ | [[File:0024-3.png|400px|thumb|left|Fig. The fragments of the constructed plasmid were recovered]] | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <br><br><br> | ||
+ | <b>M: Marker </b> | ||
+ | |||
+ | <b>1-4: ADH1(terminator)</b> | ||
+ | <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> |
Latest revision as of 05:13, 7 October 2021
pGAP-CpcA-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.
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.)
M: Marker
1-4: ADH1(terminator)
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]