Team:CCU Taiwan/Basic Part

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Kozak sequence

Prepro-alpha-factor

Mature Lac1

Mature Px18

Mature Px16

BASIC PART

BBa_K2809001
Kozak sequence (Pichia pastoris)

  The Kozak sequence initiates the translation in Pichia pastoris. This Sequence is copied directly from pCore1 in reference. The start codon is not included.

BBa_K2809002
Prepro-alpha-factor (codon modified for P. pastoris)

  Prepro-alpha-factor is the secretion signal peptide from the yeast α-mating factor. This signal peptide directs expression of secretion of the recombinant protein, therefore, it allows the exportation of it. Once the recombinant protein is in the endoplasmic reticulum (ER) lumen, this secretion signal will be cleaved.

BBa_K2809010
Mature Lac1 (laccase)

  This is the codon-optimized version of laccase derived from Pinus taeda (Loblolly pine). Laccase is involved in monolignol dehydrogenation, using oxygen to oxidize their metal centers to enable catalytical phenol oxidation. It is able to produce dimers and oligomers of coniferyl alcohol, which is involved in the early stage of lignification.

Figure 1: Three resonance forms of coniferyl alcohol with radical.

Figure2: Target gene (laccase)
The signal peptide has been removed, so the sequence does NOT contain a start codon. Furthermore, it includes HA-tag for Western Blot protein detection and 6*His-tag for protein purification before the stop codon.

BBa_K2809020
Mature Px18 (peroxidase)

  These are the codon-optimized versions of peroxidase derived from Picea abies (Norway spruce). Peroxidases (Px16 and Px18) are involved in monolignol dehydrogenation, using hydrogen peroxide as a substrate. They are able to form polymers from oligomers or dimers, which contribute to lignin polymerization.

Figure3: Three form of dimers.

Figure4: Target gene (peroxidase 16)
The signal peptide has been removed, so the sequence does NOT contain a start codon. Moreover, it includes V5-tag for Western Blot protein detection and 6*His-tag for protein purification before the stop codon.

BBa_K2809030
Mature Px16 (peroxidase)

  The function of peroxidase 16 is similar to peroxidase 18.

Figure5: Target gene (peroxidase 18)
The signal peptide has been removed, so the sequence does NOT contain a start codon. Furthermore, it includes FLAG-tag for Western Blot protein detection and 6*His-tag for protein purification before the stop codon.