Difference between revisions of "Part:BBa K5136057"
 
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<br>Enables dTDP-4-dehydrorhamnose reductase activity.<br>
 
<br>Enables dTDP-4-dehydrorhamnose reductase activity.<br>
  
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===Biology===
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==Biology==
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The rfbD encodes the dTDP-4-dehydrorhamnose reductase from <i>Escherichia coli</i> DH10β. And RfbD catalyzes the terminal step of rhamnose biosynthesis pathway by converting dTDP-6-deoxy-l-lyxo-4-hexulose to the final product dTDP-l-rhamnose, which provides precursors of rhamnose for rhamnolipids biosynthesis. (1)<br>
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The rfbD encodes the dTDP-4-dehydrorhamnose reductase from <i>Escherichia coli</i> DH10β. And RfbD catalyzes the terminal step of rhamnose biosynthesis pathway by converting dTDP-6-deoxy-l-lyxo-4-hexulose to the final product dTDP-l-rhamnose, which provides precursors of rhamnose for rhamnolipids biosynthesis (1).<br>
  
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===Usage===
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==Usage==
 
We used <partinfo>BBa_K081005</partinfo> to construct the expression system and obtained the composite <partinfo>BBa_K5136233</partinfo>, which is assembled on the expression vector pSB3K3 by standard assembly. The constructed plasmid was transformed into <i>E. coli</i> DH5α, then the positive transformants were selected by kanamycin and confirmed by colony PCR and sequencing.<br>
 
We used <partinfo>BBa_K081005</partinfo> to construct the expression system and obtained the composite <partinfo>BBa_K5136233</partinfo>, which is assembled on the expression vector pSB3K3 by standard assembly. The constructed plasmid was transformed into <i>E. coli</i> DH5α, then the positive transformants were selected by kanamycin and confirmed by colony PCR and sequencing.<br>
  
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===Characterization===
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==Characterization==
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====1. Agarose Gel Electrophoresis====
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====Agarose Gel Electrophoresis====
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After transferring the plasmid into <i>E. coli</i> DH5α, colony PCR was used to certify the plasmid was correct. The expected bands(1414 bp) were obtained at the position around 1500 bp (Fig. 1).<br>
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After transferring the plasmid into <i>E. coli</i> DH5α, colony PCR was used to certify the plasmid was correct. The expected bands(1414 bp) were obtained at the position around 1500 bp (Figure 1).<br>
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<center><html><img src="https://static.igem.wiki/teams/5136/part/yyf/j23100-rfbd.png"></html></center>
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<center><html><img src="https://static.igem.wiki/teams/5136/part/yyf/j23100-rfbd.png" width="200px"></html></center>
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<br><center><b>Fig.1 The result of colony PCR products of BBa_5136233_pSB3K3.</b></center><br>
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<br><center><b>Figure 1 The result of colony PCR products of BBa_5136233_pSB3K3.</b></center><br>
  
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===Reference===
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==Reference==
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1.Cabrera-Valladares, N. et al. (2006). "Monorhamnolipids and 3-(3-hydroxyalkanoyloxy) alkanoic acids (HAAs) production using Escherichia coli as a heterologous host." Appl. Microbiol. Biotechnol. 73(1): 187-194.<br>
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1. Cabrera-Valladares, N. et al. (2006). "Monorhamnolipids and 3-(3-hydroxyalkanoyloxy) alkanoic acids (HAAs) production using Escherichia coli as a heterologous host." Appl. Microbiol. Biotechnol. 73(1): 187-194.<br>
  
  

Latest revision as of 22:21, 1 October 2024


rfbD
Enables dTDP-4-dehydrorhamnose reductase activity.

Biology

The rfbD encodes the dTDP-4-dehydrorhamnose reductase from Escherichia coli DH10β. And RfbD catalyzes the terminal step of rhamnose biosynthesis pathway by converting dTDP-6-deoxy-l-lyxo-4-hexulose to the final product dTDP-l-rhamnose, which provides precursors of rhamnose for rhamnolipids biosynthesis (1).

Usage

We used BBa_K081005 to construct the expression system and obtained the composite BBa_K5136233, which is assembled on the expression vector pSB3K3 by standard assembly. The constructed plasmid was transformed into E. coli DH5α, then the positive transformants were selected by kanamycin and confirmed by colony PCR and sequencing.

Characterization

Agarose Gel Electrophoresis

After transferring the plasmid into E. coli DH5α, colony PCR was used to certify the plasmid was correct. The expected bands(1414 bp) were obtained at the position around 1500 bp (Figure 1).


Figure 1 The result of colony PCR products of BBa_5136233_pSB3K3.

Reference

1. Cabrera-Valladares, N. et al. (2006). "Monorhamnolipids and 3-(3-hydroxyalkanoyloxy) alkanoic acids (HAAs) production using Escherichia coli as a heterologous host." Appl. Microbiol. Biotechnol. 73(1): 187-194.


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]