Difference between revisions of "Part:BBa K2308003"
 
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<partinfo>BBa_K2308003 short</partinfo>
 
<partinfo>BBa_K2308003 short</partinfo>
  
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This part is a coding gene of sYFP2 (super yellow fluorescent protein), and the codes are optimized for expression in Rhodobacter Sphaeroides.
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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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<!-- Add more about the biology of this part here
 
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===Usage and Biology===
 
  
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2308003 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2308003 SequenceAndFeatures</partinfo>
  
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<!-- Uncomment this to enable Functional Parameter display
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===Functional Parameters===
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<partinfo>BBa_K2308003 parameters</partinfo>
 
<partinfo>BBa_K2308003 parameters</partinfo>
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<html>
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<body>
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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>
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<table class="tg">
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  <tr>
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    <th class="tg-031e">Before Optimization</th>
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    <th class="tg-yw4l">After Optimization</th>
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  </tr>
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  <tr>
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    <td class="tg-yw4l">
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ATGGTTAGCAAGGGCGAAGAACTTTTTACAGGCGTAGTACCGATCTTAGTTG
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AATTAGACGGCGACGTTAACGGTCATAAGTTTAGCGTGAGCGGTGAGGGTG
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AAGGTGACGCAACTTACGGCAAGCTGACCCTGAAGCTGATTTGCACGACGG
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GTAAGCTGCCGGTCCCGTGGCCTACCCTGGTCACGACCTTGGGTTATGGCG
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TTCAGTGTTTCGCGCGTTATCCGGACCACATGAAACAACACGATTTCTTTAA
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GAGCGCGATGCCAGAAGGCTATGTGCAGGAGCGTACGATCTTTTTCAAAGA
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CGACGGTAACTACAAGACGCGTGCCGAAGTCAAATTCGAAGGCGACACCCT
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GGTGAATCGCATTGAGCTGAAGGGTATTGATTTCAAAGAGGATGGCAATATC
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CTGGGTCACAAGCTGGAGTACAATTACAATTCCCACAACGTTTACATCACCG
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CAGATAAACAGAAAAATGGCATCAAAGCGAATTTCAAAATCCGTCACAACAT
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TGAGGACGGTGGTGTTCAACTGGCGGATCATTACCAGCAAAACACCCCGAT
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TGGTGACGGTCCGGTCCTGTTGCCGGATAACCATTATCTGTCTTACCAAAG
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CAAACTGAGCAAAGATCCGAACGAGAAGCGCGACCACATGGTGCTGCTGG
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AGTTTGTGACCGCTGCCGGTATTACCCTGGGTATGGATGAGCTGTATAAATA
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ATAA</td>
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    <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>
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  </tr>
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</table><br>
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<div class="row" align="left" style="margin-left:20%;">
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<img src="https://static.igem.org/mediawiki/2017/4/47/F1-D.png" style="height: 400px;">
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<img src="https://static.igem.org/mediawiki/2017/4/47/F1-D.png" style="height: 400px;">
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<p>Figure 1：PCR test of BBa_K2308003(optimized sYFP2) and original sYFP2</p><br><br>
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</div>
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<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>
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<div class="row" align="left" style="margin-left:10%;">
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<img src="https://static.igem.org/mediawiki/2017/b/be/Cwj_yingguang0002.jpg" style="width: 400px;">
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<img src="https://static.igem.org/mediawiki/2017/5/57/Cwj_yingguang0004.jpg" style="width: 400px;">
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<center><p>Figure 2：fluorescent image of sYFP2 (original) and sYFP2(optimized)</p></center><br><br>
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</div>
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<div class="row" align="left" style="margin-left:10%;">
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<img src="https://static.igem.org/mediawiki/2017/9/99/F5-A%281%29.png" style="width: 400px;">
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<img src="https://static.igem.org/mediawiki/2017/9/99/F5-A%281%29.png" style="width: 400px;">
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<center><p>Figure 3：Fluorescence intensity of WT, sYFP2(optimized), sYFP2(unoptimized) strains.</p></center><br><br>
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</div>
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<p> It is obvious that after the codon optimization, sYFP2 can be better expressed in <i>Rhodobacter sphaeroides 2.4.1</i>.</p><br>
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</body>
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</html>

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


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]



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

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）.


Figure 2：fluorescent image of sYFP2 (original) and sYFP2(optimized)



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.