Part:BBa_K2959005:Design

Expressible Pinus sylvestris Defensin 1 with Erv1p

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 580
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

The sequence of PsDef1 is preceded by a 6x His-Tag to facilitate its purification by immobilized metal affinity chromatography. The sequence for Erv1p consists in a truncated version that lacks the first 72 amino acids of the complete sequence. It has been proved that this version shows similar or even improved sulfhydryl oxidase activity¹.

Source

The composite part was designed using the following parts previously added to the Registry: BBa_R0010, BBa_B0034, and BBa_B0015. The amino acid sequence for PsDef1 was retrieved from Kovalyova et al. (2007). The full amino acid sequence for Erv1p was reported by Lisowsky (1996). The truncated version was retrieved from Lee et al. (2000).

References

1. Lee, J. E., Hofhaus, G., & Lisowsky, T. (2000). Erv1p from Saccharomyces cerevisiae is a FAD‐linked sulfhydryl oxidase. FEBS letters, 477(1-2), 62-66. doi: 10.1016/s0014-5793(00)01767-1
2. Lisowsky, T. (1996). Removal of an intron with unique 3′ branch site creates an amino‐terminal protein sequence directing the scERV1 gene product to mitochondria. Yeast, 12(15), 1501-1510. doi: 10.1002/(SICI)1097-0061(199612)12:15%3C1501::AID-YEA40%3E3.0.CO;2-H
3. Kovalyova, V. A., Gout, I. T., Kyamova, R. G., Filonenko, V. V., & Gout, R. T. (2007). Cloning and analysis of defensin 1 cDNA from Scots pine. Biopolymers and Cell,23(5), 398-404. doi: http://dx.doi.org/10.7124/bc.000779