Difference between revisions of "Part:BBa K2308003"
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<partinfo>BBa_K2308003 short</partinfo> | <partinfo>BBa_K2308003 short</partinfo> | ||
− | This part is a coding gene of sYFP2 (super yellow fluorescent protein), and the codes are optimized for expression in Rhodobacter | + | This part is a coding gene of sYFP2 (super yellow fluorescent protein), and the codes are optimized for expression in <i>Rhodobacter sphaeroides 2.4.1</i>. |
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<partinfo>BBa_K2308003 SequenceAndFeatures</partinfo> | <partinfo>BBa_K2308003 SequenceAndFeatures</partinfo> | ||
+ | <!-- | ||
<partinfo>BBa_K2308003 parameters</partinfo> | <partinfo>BBa_K2308003 parameters</partinfo> | ||
− | + | --> | |
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<p>sYFP2(BBa_K2308003) is the key part in our experiment. It is used both for fusion expression (see BBa_K2308002) and inducible cytoplasm expression (combined with BBa_K2308016), and the host cell is <i>Rhodobacter sphaeroides 2.4.1. </i></p><br> | <p>sYFP2(BBa_K2308003) is the key part in our experiment. It is used both for fusion expression (see BBa_K2308002) and inducible cytoplasm expression (combined with BBa_K2308016), and the host cell is <i>Rhodobacter sphaeroides 2.4.1. </i></p><br> | ||
− | + | <table class="tg"> | |
+ | <tr> | ||
+ | <th class="tg-031e">Before Optimization</th> | ||
+ | <th class="tg-yw4l">After Optimization</th> | ||
+ | </tr> | ||
+ | <tr> | ||
+ | <td class="tg-yw4l"> | ||
+ | ATGGTTAGCAAGGGCGAAGAACTTTTTACAGGCGTAGTACCGATCTTAGTTG | ||
+ | AATTAGACGGCGACGTTAACGGTCATAAGTTTAGCGTGAGCGGTGAGGGTG | ||
+ | AAGGTGACGCAACTTACGGCAAGCTGACCCTGAAGCTGATTTGCACGACGG | ||
+ | GTAAGCTGCCGGTCCCGTGGCCTACCCTGGTCACGACCTTGGGTTATGGCG | ||
+ | TTCAGTGTTTCGCGCGTTATCCGGACCACATGAAACAACACGATTTCTTTAA | ||
+ | GAGCGCGATGCCAGAAGGCTATGTGCAGGAGCGTACGATCTTTTTCAAAGA | ||
+ | CGACGGTAACTACAAGACGCGTGCCGAAGTCAAATTCGAAGGCGACACCCT | ||
+ | GGTGAATCGCATTGAGCTGAAGGGTATTGATTTCAAAGAGGATGGCAATATC | ||
+ | CTGGGTCACAAGCTGGAGTACAATTACAATTCCCACAACGTTTACATCACCG | ||
+ | CAGATAAACAGAAAAATGGCATCAAAGCGAATTTCAAAATCCGTCACAACAT | ||
+ | TGAGGACGGTGGTGTTCAACTGGCGGATCATTACCAGCAAAACACCCCGAT | ||
+ | TGGTGACGGTCCGGTCCTGTTGCCGGATAACCATTATCTGTCTTACCAAAG | ||
+ | CAAACTGAGCAAAGATCCGAACGAGAAGCGCGACCACATGGTGCTGCTGG | ||
+ | AGTTTGTGACCGCTGCCGGTATTACCCTGGGTATGGATGAGCTGTATAAATA | ||
+ | ATAA</td> | ||
+ | <td class="tg-yw4l">ATGGTGTCCAAGGGCGAGGAGGACAACATGGCCATCATCAAG<br>GAGTTCATGCGCTTCAAGGTGCACATGGAGGGCAGCGTGAACGG<br>CCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCT<br>ACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGCGGC<br>CCCCTGCCCTTCGCCTGGGACATCCTGAGCCCCCAGTTCATGTA<br>CGGCAGCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTAC<br>CTGAAGCTGAGCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGAT<br>GAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACAGCAGC<br>CTCCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACC<br>AACTTCCCCAGCGACGGCCCCGTGATGCAGAAGAAGACCATGGGCTG<br>GAGGCCAGCAGCGAGCGCATGTACCCCGAGGACGGCGCCCTGAAG<br>GGCGAGATCAAGCAGCGCCTGAAGCTGAAGGACGGCGGCCACTAC<br>GACGCCGAGGTGAAGACCACCTACAAGGCCAAGAAGCCCGTGC<br>AGCTGCCCGGCGCCTACAACGTGAACATCAAGCTGGACATCACCAGC<br>CACAACGAGGACTACACCATCGTGGAGCAGTACGAGCGCGCTG<br>AGGGCCGCCACAGCACCGGCGGCATGGACGAGCTGTACAAGTAA</td> | ||
+ | </tr> | ||
+ | </table><br> | ||
<div class="row" align="left" style="margin-left:20%;"> | <div class="row" align="left" style="margin-left:20%;"> | ||
<img src="https://static.igem.org/mediawiki/2017/4/47/F1-D.png" style="height: 400px;"> | <img src="https://static.igem.org/mediawiki/2017/4/47/F1-D.png" style="height: 400px;"> | ||
<img src="https://static.igem.org/mediawiki/2017/4/47/F1-D.png" style="height: 400px;"> | <img src="https://static.igem.org/mediawiki/2017/4/47/F1-D.png" style="height: 400px;"> | ||
− | + | <p>Figure 1:PCR test of BBa_K2308003(optimized sYFP2) and original sYFP2</p><br><br> | |
</div> | </div> | ||
− | <p> In inducible cytoplasm expression experiment, we used part BBa_K2308016 to turn the Rhodobacter | + | <p> In inducible cytoplasm expression experiment, we used part BBa_K2308016 to turn the <i>Rhodobacter sphaeroides 2.4.1</i> into inducible strains, and IPTG was added (800 μM in final volume)when the OD<sub>700</sub> of the strain was about 0.4(grown for about 24h).</p><br> |
<div class="row" align="left" style="margin-left:10%;"> | <div class="row" align="left" style="margin-left:10%;"> | ||
− | <img src="https://static.igem.org/mediawiki/2017/ | + | <img src="https://static.igem.org/mediawiki/2017/b/be/Cwj_yingguang0002.jpg" style="width: 400px;"> |
− | <img src="https://static.igem.org/mediawiki/2017/ | + | <img src="https://static.igem.org/mediawiki/2017/5/57/Cwj_yingguang0004.jpg" style="width: 400px;"> |
<center><p>Figure 2:fluorescent image of sYFP2 (original) and sYFP2(optimized)</p></center><br><br> | <center><p>Figure 2:fluorescent image of sYFP2 (original) and sYFP2(optimized)</p></center><br><br> | ||
</div> | </div> | ||
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− | <p> It is obvious that after the codon optimization, sYFP2 can be better expressed in Rhodobacter sphaeroides 2.4.1.</p><br> | + | <p> It is obvious that after the codon optimization, sYFP2 can be better expressed in <i>Rhodobacter sphaeroides 2.4.1</i>.</p><br> |
Latest revision as of 07:35, 1 November 2017
coding sequence of sYFP2 (codon optimized for Rhodobacter sphaeroides 2.4.1)
This part is a coding gene of sYFP2 (super yellow fluorescent protein), and the codes are optimized for expression in Rhodobacter sphaeroides 2.4.1.
This part is an improvement of part (BBa_K864100)
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]
sYFP2(BBa_K2308003) is the key part in our experiment. It is used both for fusion expression (see BBa_K2308002) and inducible cytoplasm expression (combined with BBa_K2308016), and the host cell is Rhodobacter sphaeroides 2.4.1.
Before Optimization | After Optimization |
---|---|
ATGGTTAGCAAGGGCGAAGAACTTTTTACAGGCGTAGTACCGATCTTAGTTG AATTAGACGGCGACGTTAACGGTCATAAGTTTAGCGTGAGCGGTGAGGGTG AAGGTGACGCAACTTACGGCAAGCTGACCCTGAAGCTGATTTGCACGACGG GTAAGCTGCCGGTCCCGTGGCCTACCCTGGTCACGACCTTGGGTTATGGCG TTCAGTGTTTCGCGCGTTATCCGGACCACATGAAACAACACGATTTCTTTAA GAGCGCGATGCCAGAAGGCTATGTGCAGGAGCGTACGATCTTTTTCAAAGA CGACGGTAACTACAAGACGCGTGCCGAAGTCAAATTCGAAGGCGACACCCT GGTGAATCGCATTGAGCTGAAGGGTATTGATTTCAAAGAGGATGGCAATATC CTGGGTCACAAGCTGGAGTACAATTACAATTCCCACAACGTTTACATCACCG CAGATAAACAGAAAAATGGCATCAAAGCGAATTTCAAAATCCGTCACAACAT TGAGGACGGTGGTGTTCAACTGGCGGATCATTACCAGCAAAACACCCCGAT TGGTGACGGTCCGGTCCTGTTGCCGGATAACCATTATCTGTCTTACCAAAG CAAACTGAGCAAAGATCCGAACGAGAAGCGCGACCACATGGTGCTGCTGG AGTTTGTGACCGCTGCCGGTATTACCCTGGGTATGGATGAGCTGTATAAATA ATAA | ATGGTGTCCAAGGGCGAGGAGGACAACATGGCCATCATCAAG GAGTTCATGCGCTTCAAGGTGCACATGGAGGGCAGCGTGAACGG CCACGAGTTCGAGATCGAGGGCGAGGGCGAGGGCCGCCCCT ACGAGGGCACCCAGACCGCCAAGCTGAAGGTGACCAAGGGCGGC CCCCTGCCCTTCGCCTGGGACATCCTGAGCCCCCAGTTCATGTA CGGCAGCAAGGCCTACGTGAAGCACCCCGCCGACATCCCCGACTAC CTGAAGCTGAGCTTCCCCGAGGGCTTCAAGTGGGAGCGCGTGAT GAACTTCGAGGACGGCGGCGTGGTGACCGTGACCCAGGACAGCAGC CTCCAGGACGGCGAGTTCATCTACAAGGTGAAGCTGCGCGGCACC AACTTCCCCAGCGACGGCCCCGTGATGCAGAAGAAGACCATGGGCTG GAGGCCAGCAGCGAGCGCATGTACCCCGAGGACGGCGCCCTGAAG GGCGAGATCAAGCAGCGCCTGAAGCTGAAGGACGGCGGCCACTAC GACGCCGAGGTGAAGACCACCTACAAGGCCAAGAAGCCCGTGC AGCTGCCCGGCGCCTACAACGTGAACATCAAGCTGGACATCACCAGC CACAACGAGGACTACACCATCGTGGAGCAGTACGAGCGCGCTG AGGGCCGCCACAGCACCGGCGGCATGGACGAGCTGTACAAGTAA |
![](https://static.igem.org/mediawiki/2017/4/47/F1-D.png)
![](https://static.igem.org/mediawiki/2017/4/47/F1-D.png)
Figure 1:PCR test of BBa_K2308003(optimized sYFP2) and original sYFP2
In inducible cytoplasm expression experiment, we used part BBa_K2308016 to turn the Rhodobacter sphaeroides 2.4.1 into inducible strains, and IPTG was added (800 μM in final volume)when the OD700 of the strain was about 0.4(grown for about 24h).
![](https://static.igem.org/mediawiki/2017/b/be/Cwj_yingguang0002.jpg)
![](https://static.igem.org/mediawiki/2017/5/57/Cwj_yingguang0004.jpg)
Figure 2:fluorescent image of sYFP2 (original) and sYFP2(optimized)
![](https://static.igem.org/mediawiki/2017/9/99/F5-A%281%29.png)
![](https://static.igem.org/mediawiki/2017/9/99/F5-A%281%29.png)
Figure 3:Fluorescence intensity of WT, sYFP2(optimized), sYFP2(unoptimized) strains.
It is obvious that after the codon optimization, sYFP2 can be better expressed in Rhodobacter sphaeroides 2.4.1.