Difference between revisions of "Part:BBa K5136058"
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<br>Required for rhamnolipid surfactant production. | <br>Required for rhamnolipid surfactant production. | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
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===<br><b>Biology</b><br>=== | ===<br><b>Biology</b><br>=== | ||
<br>The RhlA comes from <i>Pseudomonas aeruginosa</i> and can compete with the enzymes of the type II fatty acid synthase (FASII) cycle for the β-hydroxyacyl-acyl carrier protein (ACP) pathway intermediates, therefore supplies the acyl moieties for rhamnolipid biosynthesis. RhlA has a greater affinity for 10-carbon substrates, and is the only enzyme required to generate the lipid component of rhamnolipid. (1)<br> | <br>The RhlA comes from <i>Pseudomonas aeruginosa</i> and can compete with the enzymes of the type II fatty acid synthase (FASII) cycle for the β-hydroxyacyl-acyl carrier protein (ACP) pathway intermediates, therefore supplies the acyl moieties for rhamnolipid biosynthesis. RhlA has a greater affinity for 10-carbon substrates, and is the only enzyme required to generate the lipid component of rhamnolipid. (1)<br> | ||
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===<br><b>Usage</b><br>=== | ===<br><b>Usage</b><br>=== | ||
<br>We used <partinfo>BBa_K081005</partinfo> to construct the expression system and obtained the composite <partinfo>BBa_K5136232</partinfo>, which is assembled on the expression vector pSB1C3 by standard assembly. The constructed plasmid was transformed into <i>E. coli</i> DH5α, then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing.<br> | <br>We used <partinfo>BBa_K081005</partinfo> to construct the expression system and obtained the composite <partinfo>BBa_K5136232</partinfo>, which is assembled on the expression vector pSB1C3 by standard assembly. The constructed plasmid was transformed into <i>E. coli</i> DH5α, then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing.<br> | ||
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===<br><b>Characterization</b><br>=== | ===<br><b>Characterization</b><br>=== | ||
====<br><b>1. Agarose Gel Electrophoresis</b><br>==== | ====<br><b>1. Agarose Gel Electrophoresis</b><br>==== | ||
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<br>Fig.1 The result of colony PCR products of <partinfo>BBa_K5136232</partinfo>_pSB1C3.<br> | <br>Fig.1 The result of colony PCR products of <partinfo>BBa_K5136232</partinfo>_pSB1C3.<br> | ||
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===<br><b>Reference</b><br>=== | ===<br><b>Reference</b><br>=== | ||
<br>1. Zhu, K.; Rock, C. O., RhlA converts beta-hydroxyacyl-acyl carrier protein intermediates in fatty acid synthesis to the beta-hydroxydecanoyl-beta-hydroxydecanoate component of rhamnolipids in Pseudomonas aeruginosa. J Bacteriol 2008, 190 (9), 3147-54.<br> | <br>1. Zhu, K.; Rock, C. O., RhlA converts beta-hydroxyacyl-acyl carrier protein intermediates in fatty acid synthesis to the beta-hydroxydecanoyl-beta-hydroxydecanoate component of rhamnolipids in Pseudomonas aeruginosa. J Bacteriol 2008, 190 (9), 3147-54.<br> | ||
Revision as of 07:07, 17 September 2024
rhlA
Required for rhamnolipid surfactant production.
Biology
The RhlA comes from Pseudomonas aeruginosa and can compete with the enzymes of the type II fatty acid synthase (FASII) cycle for the β-hydroxyacyl-acyl carrier protein (ACP) pathway intermediates, therefore supplies the acyl moieties for rhamnolipid biosynthesis. RhlA has a greater affinity for 10-carbon substrates, and is the only enzyme required to generate the lipid component of rhamnolipid. (1)
Usage
We used BBa_K081005 to construct the expression system and obtained the composite BBa_K5136232, which is assembled on the expression vector pSB1C3 by standard assembly. The constructed plasmid was transformed into E. coli DH5α, then the positive transformants were selected by chloramphenicol and confirmed by colony PCR and sequencing.
Characterization
1. Agarose Gel Electrophoresis
After transferring the plasmid into E. coli DH5α, colony PCR was used to certify the plasmid was correct. The expected bands (bp) was obtained at the position around bp (Fig. 1).
Fig.1 The result of colony PCR products of BBa_K5136232_pSB1C3.
Reference
1. Zhu, K.; Rock, C. O., RhlA converts beta-hydroxyacyl-acyl carrier protein intermediates in fatty acid synthesis to the beta-hydroxydecanoyl-beta-hydroxydecanoate component of rhamnolipids in Pseudomonas aeruginosa. J Bacteriol 2008, 190 (9), 3147-54.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 69
Illegal XhoI site found at 805 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 294
Illegal BsaI.rc site found at 478