Moreover, the Bam HI site between the AOX1
promoter and signal was deleted so that the promoter is linked with the signal peptide directly.
coli (BL21) pLysS LB + 37,20 Bnl2 and pET-26(+) antibiotics Thermomyces Pichia GS115 and BMGY lanuginosus (lipase expression plasmid medium 28,28 gene lnl) vector pPIC9K (Eleterologous expression under AOX1
promoter) Geobacillus E.
The vector contains the alcohol oxidase (aox1
) promoter from P.
Ten micrograms of linearized, recombinant plasmid DNA was used to transform the KM71H (aox1
promoter has been utilized for production of heterologous protein in P.pastoris.
The inducible promoter from AOX1
gene, the chemical synthesized signal MF4I, the native transcription termination and polyadenylation signal from AOX1
gene consist pYPX8 (16).
pastoris under the control of the AOX1
promoter with the a-factor signal sequence for secretion.
Pichia pasloris.A PCR-amplified DNA fragment from the CUZD1 cDNA transcript variant 1 (Origene) encoding the 3' His-tagged extracellular domain of CUZD1 isoform 1 (amino acids 1-568 of NCBI GenBank accession number NP_071317), flanked by XhoI and NotI restriction enzyme sites, was inserted into the multiple cloning site of the pPIC9 yeast expression vector, downstream from the Pichia [alpha]-secretion signal and the aldehyde oxidase 1 (AOX1
P pastoris alcohol oxidase has two isoforms which are the products of AOX1
and AOX2 genes.
For induction of AOX1
promoter and protein production, the growth medium was transferred to the same medium except with 0.5% methanol in place with glycerol as the carbon source .