Difference between revisions of "Part:BBa K2308003"

Line 31: Line 31:
 
   <tr>
 
   <tr>
 
     <td class="tg-yw4l">
 
     <td class="tg-yw4l">
ATGGTTAGCAAGGGCGAAGAACTTTTTACAGGCGTAGTACCGATCTTAGTTGAATTAGACGGCGACGTTAACGGTCATAAGTTTAGCGTGAGCGGTGAGG
+
ATGGTTAGCAAGGGCGAAGAACTTTTTACAGGCGTAGTACCGATCTTAGTTG
GTGAAGGTGACGCAACTTACGGCAAGCTGACCCTGAAGCTGATTTGCACGACGGGTAAGCTGCCGGTCCCGTGGCCTACCCTGGTCACGACCTTGGGTTA
+
AATTAGACGGCGACGTTAACGGTCATAAGTTTAGCGTGAGCGGTGAGGGTG
TGGCGTTCAGTGTTTCGCGCGTTATCCGGACCACATGAAACAACACGATTTCTTTAAGAGCGCGATGCCAGAAGGCTATGTGCAGGAGCGTACGATCTTT
+
AAGGTGACGCAACTTACGGCAAGCTGACCCTGAAGCTGATTTGCACGACGG
TTCAAAGACGACGGTAACTACAAGACGCGTGCCGAAGTCAAATTCGAAGGCGACACCCTGGTGAATCGCATTGAGCTGAAGGGTATTGATTTCAAAGAGG
+
GTAAGCTGCCGGTCCCGTGGCCTACCCTGGTCACGACCTTGGGTTATGGCG
ATGGCAATATCCTGGGTCACAAGCTGGAGTACAATTACAATTCCCACAACGTTTACATCACCGCAGATAAACAGAAAAATGGCATCAAAGCGAATTTCAA
+
TTCAGTGTTTCGCGCGTTATCCGGACCACATGAAACAACACGATTTCTTTAA
AATCCGTCACAACATTGAGGACGGTGGTGTTCAACTGGCGGATCATTACCAGCAAAACACCCCGATTGGTGACGGTCCGGTCCTGTTGCCGGATAACCAT
+
GAGCGCGATGCCAGAAGGCTATGTGCAGGAGCGTACGATCTTTTTCAAAGA
TATCTGTCTTACCAAAGCAAACTGAGCAAAGATCCGAACGAGAAGCGCGACCACATGGTGCTGCTGGAGTTTGTGACCGCTGCCGGTATTACCCTGGGTA
+
CGACGGTAACTACAAGACGCGTGCCGAAGTCAAATTCGAAGGCGACACCCT
TGGATGAGCTGTATAAATAATAA</td>
+
GGTGAATCGCATTGAGCTGAAGGGTATTGATTTCAAAGAGGATGGCAATATC
     <td class="tg-yw4l">ATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAGCTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGATGCCACCTACGGCAAGCTGACCCTGAAGCTGATCTGCACCACCGGCAAGCTGCCCGTGCCCTGGCCCACCCTCGTGACCACCCTGGGCTACGGCGTGCAGTGCTTCGCCCGCTACCCCGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAGGAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGTTCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGGACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATCACCGCCGACAAGCAGAAGAACGGCATCAAGGCCAACTTCAAGATCCGCCACAACATCGAGGACGGCGGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACGGCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCTACCAGTCCAAGCTGAGCAAAGACCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGATCACTCTCGGCATGGACGAGCTGTACAAGTAATGA</td>
+
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>
 
   </tr>
 
</table><br>
 
</table><br>

Revision as of 11:02, 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:
  • 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]
directionforward



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 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.