Difference between revisions of "Part:BBa K3739053"
 
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===Usage===
 
===Usage===
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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 composite parts BBa_K3739046 and BBa_K3739047 together into the expression vector pET-28a(+) by standard assembly, then transferred it into ''Vibrio natriegens'' to verify its ability to produce rhamnolipids.
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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 composite parts <partinfo>BBa_K3739046</partinfo> and <partinfo>BBa_K3739047</partinfo> together into the expression vector pET-28a(+) by standard assembly, then transferred it into ''Vibrio natriegens'' to verify its ability to produce rhamnolipids.
  
 
===Characterization===
 
===Characterization===
  
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1. Agarose Gel Electrophoresis
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=====1. Agarose Gel Electrophoresis(rhlA)=====
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After transferring the plasmid into ''Vibrio natriegens'', regular PCR was used to certify the plasmid was correct. The expected result was obtained(2821bp).  
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After transferring the plasmid into ''Vibrio natriegens'', colony PCR was used to certify the plasmid was correct. The expected result was obtained(1305bp).<br/>
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[[File:T--XMU-China--12F1-1.png|300px]]<br/>
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'''Fig.1''' The result of colony PCR. Plasmid pET-28a(+).
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=====2. Agarose Gel Electrophoresis(rhlB)=====
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After receiving the plasmid from Sangon Biotech®, regular PCR was used to certify the correctness of the plasmid. Target bands (1698 bp) can be observed at the position around 1700 bp (Fig. 2).<br/>
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[[File:T--XMU-China--13-F1.png|300px]]<br/>
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'''Fig.2''' The result of regular PCR. Plasmid pET-28a(+).
  
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Fig.1 The result of regular PCR. Plasmid pET-28a(+).
 
  
  
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===Reference===
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1. Chong, H.; Li, Q., Microbial production of rhamnolipids: opportunities, challenges and strategies. ''Microb Cell Fact'' '''2017''', 16 (1), 137.
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1. Chong, H.; Li, Q., Microbial production of rhamnolipids: opportunities, challenges and strategies. Microb Cell Fact 2017, 16 (1), 137.
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Latest revision as of 00:16, 22 October 2021


J23100-B0034-his-rhlA-B0015-J23100-B0034-rhlB-B0015

Responsible for bioproduction of rhamnolipid.

Biology

RhlAB

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

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 composite parts BBa_K3739046 and BBa_K3739047 together into the expression vector pET-28a(+) by standard assembly, then transferred it into Vibrio natriegens to verify its ability to produce rhamnolipids.

Characterization

1. Agarose Gel Electrophoresis(rhlA)

After transferring the plasmid into Vibrio natriegens, colony PCR was used to certify the plasmid was correct. The expected result was obtained(1305bp).
T--XMU-China--12F1-1.png
Fig.1 The result of colony PCR. Plasmid pET-28a(+).

2. Agarose Gel Electrophoresis(rhlB)

After receiving the plasmid from Sangon Biotech®, regular PCR was used to certify the correctness of the plasmid. Target bands (1698 bp) can be observed at the position around 1700 bp (Fig. 2).
T--XMU-China--13-F1.png
Fig.2 The result of regular PCR. Plasmid pET-28a(+).


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 7
    Illegal NheI site found at 30
    Illegal NheI site found at 1121
    Illegal NheI site found at 1144
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 150
    Illegal XhoI site found at 886
    Illegal XhoI site found at 2333
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1226
    Illegal NgoMIV site found at 2060
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 375
    Illegal BsaI site found at 1576
    Illegal BsaI.rc site found at 559