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| | ===Usage and Biology=== | | ===Usage and Biology=== |
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| − | <p style="font-size: 180%; font-weight: bold;">Build:
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| − | <p style="font-size: 160%; font-weight: bold;">1.Experimental Methods
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| − | <p>1.1 Construction of PCWJ-YciA-sfp-MmCAR-YahK<br>
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| − | The pCWJ plasmid was extracted from a laboratory conserved strain. In-Fusion cloning was used to ligate the above vector to the Ptrc-YciA-sfp-MmCAR-YahK fragment.
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| − | <br>1.2 Construction of PRSFDuet-YciA-sfp-MmCAR-YahK<br>
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| − | The PRSFDuet plasmid was extracted from laboratory conserved strains. The fragment Ptrc-YciA-sfp-MmCAR-sfp-Yahk was amplified by PCR using PCWJ-YciA-sfp-MmCAR-YahK as a template, and Ptrc-YciA-MmCAR-sfp-Yahk was ligated by In-fusion cloning to the vector PRSFDuet that was linearized using PCR for homologous recombination ligation.
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| − | <p style="font-size: 160%; font-weight: bold;">Result
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| − | <p>By comparing the fermentation production of 1,5-PDO content with the previous KA30 with PCWJ+PTRC99a, we can clearly see that KA30 with PRSFDuet+PTRC99a produced more 1,5-PDO, therefore, we decided to use both vectors, PRSFDuet and PTRC99a, in all our future experiments. </p >
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Revision as of 21:57, 11 October 2023
Ptrc-sfp-MmCAR-YahK
(1)Ptrc is a promoter.(2)YahK is a acyl-CoA thioester hydrolase from Haemophilus influenzae.(3) Sfp is a phosphopantetheinyl transferase from Bacilus subtilis.(4) MmCAR is a Carboxylic acid reductase from Mycobacterium marinum.(5) YahK is a NADPH-dependent aldehyde reductase.
