Difference between revisions of "Part:BBa K5136232"
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===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 constant promoter <partinfo>BBa_J23100</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, then transferred it into E. coli BL21(DE3) to verify its ability to produce rhamnolipids.<br>
  
 
===Characterization===
 
===Characterization===

Revision as of 07:48, 17 September 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 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.

Characterization

1. Double transformation

We used double transformation to prove the monorhamnoolipid production. Plasmid BBa_K5136233_pSB3K3 and plasmid BBa_K5136232_pSB1C3 were transformed into E. coli BL21(DE3). The positive transformants were selected by kanamycin and chloramphenicol.

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