Difference between revisions of "Part:BBa K2308003"
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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"> | ||
| + | ATGGTTAGCAAGGGCGAAGAACTTTTTACAGGCGTAGTACCGATCTTAGTTGAATTAGACGGCGACGTTAACGGTCATAAGTTTAGCGTGAGCGGTGAGG | ||
| + | GTGAAGGTGACGCAACTTACGGCAAGCTGACCCTGAAGCTGATTTGCACGACGGGTAAGCTGCCGGTCCCGTGGCCTACCCTGGTCACGACCTTGGGTTA | ||
| + | TGGCGTTCAGTGTTTCGCGCGTTATCCGGACCACATGAAACAACACGATTTCTTTAAGAGCGCGATGCCAGAAGGCTATGTGCAGGAGCGTACGATCTTT | ||
| + | TTCAAAGACGACGGTAACTACAAGACGCGTGCCGAAGTCAAATTCGAAGGCGACACCCTGGTGAATCGCATTGAGCTGAAGGGTATTGATTTCAAAGAGG | ||
| + | ATGGCAATATCCTGGGTCACAAGCTGGAGTACAATTACAATTCCCACAACGTTTACATCACCGCAGATAAACAGAAAAATGGCATCAAAGCGAATTTCAA | ||
| + | AATCCGTCACAACATTGAGGACGGTGGTGTTCAACTGGCGGATCATTACCAGCAAAACACCCCGATTGGTGACGGTCCGGTCCTGTTGCCGGATAACCAT | ||
| + | TATCTGTCTTACCAAAGCAAACTGAGCAAAGATCCGAACGAGAAGCGCGACCACATGGTGCTGCTGGAGTTTGTGACCGCTGCCGGTATTACCCTGGGTA | ||
| + | TGGATGAGCTGTATAAATAATAA</td> | ||
| + | <td class="tg-yw4l">ATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGCTGATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGGGCTACGGCGTGCAGTGCTTCGCCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCACCGCCGACAAGCAGAAGAACGGCATCAAGGCCAACTTCAAGATCCGCCACAACATCGAGGACGGCGGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCTACCAGTCCAAGCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAATGA</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;"> | ||
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<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/b/be/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> | ||
Revision as of 10:52, 31 October 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.
This part is an improvement of part (BBa_K864100)
Sequence and Features
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
| direction | forward |
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 |
|---|---|
| ATGGTTAGCAAGGGCGAAGAACTTTTTACAGGCGTAGTACCGATCTTAGTTGAATTAGACGGCGACGTTAACGGTCATAAGTTTAGCGTGAGCGGTGAGG GTGAAGGTGACGCAACTTACGGCAAGCTGACCCTGAAGCTGATTTGCACGACGGGTAAGCTGCCGGTCCCGTGGCCTACCCTGGTCACGACCTTGGGTTA TGGCGTTCAGTGTTTCGCGCGTTATCCGGACCACATGAAACAACACGATTTCTTTAAGAGCGCGATGCCAGAAGGCTATGTGCAGGAGCGTACGATCTTT TTCAAAGACGACGGTAACTACAAGACGCGTGCCGAAGTCAAATTCGAAGGCGACACCCTGGTGAATCGCATTGAGCTGAAGGGTATTGATTTCAAAGAGG ATGGCAATATCCTGGGTCACAAGCTGGAGTACAATTACAATTCCCACAACGTTTACATCACCGCAGATAAACAGAAAAATGGCATCAAAGCGAATTTCAA AATCCGTCACAACATTGAGGACGGTGGTGTTCAACTGGCGGATCATTACCAGCAAAACACCCCGATTGGTGACGGTCCGGTCCTGTTGCCGGATAACCAT TATCTGTCTTACCAAAGCAAACTGAGCAAAGATCCGAACGAGAAGCGCGACCACATGGTGCTGCTGGAGTTTGTGACCGCTGCCGGTATTACCCTGGGTA TGGATGAGCTGTATAAATAATAA | ATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGCTGATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGGGCTACGGCGTGCAGTGCTTCGCCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCACCGCCGACAAGCAGAAGAACGGCATCAAGGCCAACTTCAAGATCCGCCACAACATCGAGGACGGCGGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCTACCAGTCCAAGCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAATGA |
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 sphaedoides 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.