Difference between revisions of "Part:BBa K1696010"
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<partinfo>BBa_K1696010 short</partinfo>
 
<partinfo>BBa_K1696010 short</partinfo>
  
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The riboflavin biosynthesis genes ribABDEC from ''E. coli'' are overexpressed under the control of the inducible trc promoter. Enzymes encoded by the genes shown are: ribA, GTP cyclohydrolase II; ribB, 3,4-dihydroxy-2-butanone 4-phosphate synthase; ribD, fused diaminohydroxyphosphoribosylaminopyrimidine deaminase/ 5-amino-6-(5-phosphoribosylamino)uracil reductase; ribE, 6,7-dimethyl-8-ribityllumazine synthase; ribC, riboflavin synthase gene.
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We construct this part based on BBa_K169011 and BBa_K1172303.
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<img src="https://static.igem.org/mediawiki/parts/c/c3/屏幕快照_2015-09-17_下午2.47.23.png" width="400px"/>
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Fig1.The design of this part.
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Based on co-factor model, EET pathway points out that FMN plays a critical role in electron transfer. Through the study by Dr. Tao Chen, they have weakened the RBS upstream of ribC to divert more of the material flux to RF production. Based on their viewpoint, we got further to design a strong RBS sequence upstream of the ribC and we rename the new part as EC10*(BBa_K1696010). The yield of EC10* reached 90 mg/L(as shown in Figure2). The strong RBS before ribC sequence can lead to the FMN accumulation while riboflavin consumes a bit, which may in turn, result in increase both for riboflavin and FMN as a whole.
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<img src="https://static.igem.org/mediawiki/2015/2/20/QQ图片20150918005538.jpg" width="400px"/>
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Figure 2. The yield of riboflavin in different strains: EC10(BBa_K1696011), Rf02S (Δpgi + EC10), EC10*(this part)
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<img src="https://static.igem.org/mediawiki/2015/2/20/QQ图片20150918005538.jpg" width="500px"/>
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Figure 3. The yield of riboflavin in different strains: EC10, Rf02S (Δpgi + EC10), EC10*
  
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We construct this part based on BBa_K1696010. We change the RBS-C to a stronger one. So both riboflavin and FMN will improve.
 
  
 
<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 11:55, 18 September 2015

Riboflavin enhancing parts

We construct this part based on BBa_K169011 and BBa_K1172303.

Fig1.The design of this part.

Based on co-factor model, EET pathway points out that FMN plays a critical role in electron transfer. Through the study by Dr. Tao Chen, they have weakened the RBS upstream of ribC to divert more of the material flux to RF production. Based on their viewpoint, we got further to design a strong RBS sequence upstream of the ribC and we rename the new part as EC10*(BBa_K1696010). The yield of EC10* reached 90 mg/L(as shown in Figure2). The strong RBS before ribC sequence can lead to the FMN accumulation while riboflavin consumes a bit, which may in turn, result in increase both for riboflavin and FMN as a whole.

Figure 2. The yield of riboflavin in different strains: EC10(BBa_K1696011), Rf02S (Δpgi + EC10), EC10*(this part)

Figure 3. The yield of riboflavin in different strains: EC10, Rf02S (Δpgi + EC10), EC10*


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3386
    Illegal BamHI site found at 1370
    Illegal XhoI site found at 64
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 262
    Illegal AgeI site found at 992
    Illegal AgeI site found at 1034
    Illegal AgeI site found at 2752
    Illegal AgeI site found at 3139
  • 1000
    COMPATIBLE WITH RFC[1000]