Difference between revisions of "Part:BBa K5136232"
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===Biology===
 
===Biology===
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RhlA, RhlB and RhlC, which are important enzymes to control the production of rhamnolipids, are found almost exclusively in <i>Pseudomonas sp.</i> and <i>Burkholderia sp.</i> (1)<br>
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RhlA, RhlB and RhlC, which are important enzymes to control the production of rhamnolipids, are found almost exclusively in <i>Pseudomonas</i> sp. and <i>Burkholderia</i> sp. (1).<br>
  
 
===Usage and Design===
 
===Usage and Design===
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RhlA is used for generating the lipid component of rhamnolipid, RhlB can then catalyze the reaction between HAA and dTDP-l-rhamnose to form mono-rhamnolipid. We ligased two basic parts, then they are co-expression by the constant promoter BBa_J23100. And the RBS BBa_B0034 as well as the terminator BBa_B0015 together into the expression vector pSB1C3 by standard assembly, then transferred it into E. coli BL21(DE3) to verify its ability to produce rhamnolipids.<br>
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RhlA is used for generating the lipid component of rhamnolipid, RhlB can then catalyze the reaction between HAA and dTDP-l-rhamnose to form mono-rhamnolipid. We ligased two basic parts, then they are co-expression by the promoter <partinfo>BBa_I0500</partinfo>. And the RBS <partinfo>BBa_B0034</partinfo> as well as the terminator <partinfo>BBa_B0015</partinfo> together into the expression vector pSB1C3 by standard assembly (Figure 1), then transferred it into <i>E. coli</i> BL21(DE3) to verify its ability to produce rhamnolipids.<br>
  
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===Characterization===
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<center><html><img src="https://static.igem.wiki/teams/5136/part/yyf/i0500-b0034-rhla-b0034-rhlb-b0015-psb1c3.jpg" width="400px"></html></center><br>
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====1. Double transformation====
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<center><b>Figure 1 Gene circuits of verification system for rhamnolipids production. </b></center>
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We used double transformation to prove the monorhamnoolipid production. Plasmid <partinfo>BBa_K5136233</partinfo>_pSB3K3 and plasmid <partinfo>BBa_K5136232</partinfo>_pSB1C3 were transformed into <i>E. coli</i> BL21(DE3). The positive transformants were selected by kanamycin and chloramphenicol.<br>
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===Reference===
 
===Reference===

Latest revision as of 07:03, 2 October 2024


I0500-B0034-rhlA-B0034-rhlB-B0015
Responsible for bioproduction of rhamnolipid.

Biology

RhlA, RhlB and RhlC, which are important enzymes to control the production of rhamnolipids, are found almost exclusively in Pseudomonas sp. and Burkholderia sp. (1).

Usage and Design

RhlA is used for generating the lipid component of rhamnolipid, RhlB can then catalyze the reaction between HAA and dTDP-l-rhamnose to form mono-rhamnolipid. We ligased two basic parts, then they are co-expression by the promoter BBa_I0500. And the RBS BBa_B0034 as well as the terminator BBa_B0015 together into the expression vector pSB1C3 by standard assembly (Figure 1), then transferred it into E. coli BL21(DE3) to verify its ability to produce rhamnolipids.


Figure 1 Gene circuits of verification system for rhamnolipids production.

Reference

1. Chong, H.; Li, Q., Microbial production of rhamnolipids: opportunities, challenges and strategies. Microb Cell Fact 2017, 16 (1), 137.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
    Illegal BamHI site found at 1305
    Illegal XhoI site found at 2041
    Illegal XhoI site found at 3288
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 2181
    Illegal NgoMIV site found at 3015
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1530
    Illegal BsaI site found at 2531
    Illegal BsaI.rc site found at 1714
    Illegal SapI site found at 961