Difference between revisions of "Part:BBa K398006:Design"

(→‎Design Notes)
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                     QVLSPTVPFGGYKQSGLGRELGMQALDAYLETKTVICDLNDRPMTLF
 
                     QVLSPTVPFGGYKQSGLGRELGMQALDAYLETKTVICDLNDRPMTLF
  
−
According to the authors [1], the purified protein has an activity of dodecanal dehydrogenase equal to 22.995 U/mg protein (4e-7 kat/mg protein).
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According to the authors [1], the purified protein has an activity of dodecanal dehydrogenase equal to 22.995 U/mg protein (4e-7 kat/mg protein). This part was expressed using the promoter-rbs combination J23100-J61117 and also J13002. Thus generating the parts: [https://parts.igem.org/Part:BBa_K398029 BBa_K398029] and [https://parts.igem.org/Part:BBa_K398030 BBa_K398030], respectively.
  
 
===Source===
 
===Source===

Revision as of 13:44, 27 October 2010

Medium-Chain Aldehyde Dehydrogenase


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

The original CDS can be found [http://www.ncbi.nlm.nih.gov/nuccore/AB047106.1 in this link], the file reports a cluster of proteins induced by alkanes. We took the ORF contained from 1213 to 2706. Nucleotide sequence optimized for expression in E.coli K12. We used [http://www.jcat.de/ jcat] for this purpose. Moreover, we deleted the restriction sites for EcoRI, PstI, SpeI and XbaI and we change the stop codon for a TAA.

The expected aminoacid sequence is: 

                    MTMISAQTTEADLWIDGEWRPAASGERFDVIDPATGEVTARVAN
                    AGEDDVDAAVAIAEEAFSVRRWLAISPLERGRILRRIAELIRQHHCELAQLMTRENGM
                    PINLALFIEIPLADCFDFFASLVVKPQGEVLPFSVAGSAPDYMAWTMKEPIGVAGLIT
                    PWNFPLLMPTWKIAPALAAGCTMVVKPAPETPLTALKLAEICHEAGVPEGVINVLPGL
                    DEAGKALVRHPRVPKIAFTGETETGRHILQAAAPHIKRVTLELGGKSPNIIFADADLE
                    QAAKSALFGVFYNSGQVCQAGSRILVERTVYEPFVERLAERAKKLKVGPGTNPRSDLG
                    PVISREQYEKVLRYIEIGKQEGARLAAGGRALDGGGGGYFIEPTVFADVSPSMRIACE
                    EIFGPVAAVIPFADEEEAVRIANGTMYGLAAAVWTNDIKRALRLARRVKSGTVWLNTY
                    QVLSPTVPFGGYKQSGLGRELGMQALDAYLETKTVICDLNDRPMTLF

According to the authors [1], the purified protein has an activity of dodecanal dehydrogenase equal to 22.995 U/mg protein (4e-7 kat/mg protein). This part was expressed using the promoter-rbs combination J23100-J61117 and also J13002. Thus generating the parts: BBa_K398029 and BBa_K398030, respectively.

Source

Genbank accession number: [http://www.ncbi.nlm.nih.gov/nuccore/AB047106 AB047106]

References

  1. Kato T. et al. "Gene cloning and characterization of an aldehyde dehydrogenase from long-chain alkane-degrading Geobacillus thermoleovorans B23.", Extremophiles. 2010 Jan;14(1):33-9. Epub 2009 Sep 29.
  2. National Center for Biotechnology Information [http://www.ncbi.nlm.nih.gov/]