Part:BBa_K2308003

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.