Difference between revisions of "Part:BBa K2308003"
| Line 3: | Line 3: | ||
<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>. |
| Line 57: | Line 57: | ||
| − | <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 OD700 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%;"> | ||
| Line 72: | Line 72: | ||
| − | <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> |
Revision as of 11:22, 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 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]
| 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 |
|---|---|
| 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.