phaffii PDI gene was re-amplified (oligos 447/448) from the in-house plasmid pPIC3.5 K-PDI [30]. The primer 448 introduces six His-codons followed by the TAA stop-codon. After excision with NotI and MfeI, the 1.64-kb DNA (PDIH) fragment was cloned into NotI and MfeI-digested pPICZ ...
The plasmids used in this study were generated from the pGAPZ_A and pPIC3.5K vectors (Invitrogen). Detailed description of plasmid construction is provided in Additional file 1. Yeast transformation was performed as previously described using an electroporator [26]. The following parameters were used...
PICXYN18 is the yeast strain transformed with the xyn11A cDNA in plasmid pPIC3.5. Control was transformed with pPIC3.5 vector alone. A) Xylan degradation halo produced on plates by the strain expressing Xyn11A. B) Xylanase activity in the culture supernatant of PICXYN18 (white square) or...
The PCR fragment was ligated into the respective sites of pPIC9K resulting in pPIC9K-proROL under the control of the methanol inducible alcohol oxidase 1 promoter (P AOX1 ) and fused in-frame with the α-factor secretion signal peptide of S. cerevisiae. For construction of the chimeric ...
The recombinant pPIC9K plasmids were linearized before transfered by electroporation into Pichia pastoris GS115. Selection of His+ transformants and G418-resistant clones was carried out according to the manufacturer's instruction manual (Invitrogen, USA). The selected clones were first cultured in 3...
pastoris X-33(pPIC-PA1) was studied in a standard, four phase process in a stirred bioreactor. To evaluate the maximum synthetic capacity of the yeast system for the intracellular expression of hPGAEc, a transient phase of the process was optimized (the phase required for the transition of ...
A different form of transmission-blocking vaccine, the RTS, S vaccine that blocks transmission of sporozoites from mosquitoes to humans, has recently demonstrated significant reduction in both uncomplicated and severe malaria in trials in Mozambique [5] and similar effects on both transmission and ...