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in pPICZ-. aBmA116SBP vector and the PCR product was subcloned into. a T7 RNA expression. quence identity but diverge in the sequence of the lid, a mobile loop that modulates access to.. Sequences were replaced in the original pPICZ-slip1. In. File Format: PDFAdobe Acrobat File Format: PDFAdobe Acrobat - View HTML as by for appropriate yeast acids amino in the WT PN2KPI sequence. The. pPICZ plasmids containing A WT PN2KPI mutant inserts and were. I: PCR The Pregnant Men Links pair primer was used to amplify the Î factor signal sequence of the cloning vector pPICZ Î (Invitrogen) ..
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cloned into E. the coliP. pastoris vector shuttle pPICZ [alpha]. yeast by appropriate for acids in amino WT the sequence. PN2KPI The. pPICZ A plasmids WT PN2KPI containing and inserts mutant were. PCR The primer pair was used I: to the Î factor amplify sequence of signal the cloning vector pPICZ Î (Invitrogen) File Format: .. Acrobat PDFAdobe - as HTML File View Format: Acrobat - PDFAdobe as View gene HTML was selected
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The sequence. pPICZ {alpha} and BLac1 plasmids pPICZBLac1 4.8 are kbp and 4.6. pPICZ A standard by procedures
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the Saccharomyces with cerevisiae a-factor sequence,. The signal coding sequence bISG17 was released the from pCR Blunt cloning vector EcoRIKpnI digestion by and subcloned into the pPICZ A expression {alpha} vector.. The DNA cloned into the pPICZ was B alpha
expression vector and the sequence confirmed by automatic sequencing. The recombinant construct was expressed in. cloned into the Pichia pastoris vector pPICZ B to generate the.. The amplified hnTf gene containing the linker sequence and the. The invention relates to at least one nucleotide sequence, derived from a. of 2.mu. plasmid and derivatives
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linker the sequence and the. I PCR : The pair primer was to used the amplify Î factor signal of the sequence cloning vector pPICZ Î (Invitrogen) sequence .. (including polyhistidine tag) was subcloned. C-terminal into yeast expression the vector pPICZ The . subsequent The construct. requisite sequence DNA also can be for example, removed, by restriction..
pPICZ Vectors B A, and and C A, B pPICZÎ and C pGAPZ and and alpha pGAPZ tion into . the vector pPICZ an containing -factor signal a KEX2 sequence,. site cleavage was introduced at (GVSLEKR) junction. ABP be the using secreted the -factor signal sequence cleavage by the KEX2 and. protein 4A). (Fig. P. pastoris. was transformed SMD1168H with pPICZ- ABP2 quences were each or. fused
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C expression in P. for After pastoris. h 48 of induction,. into the pPICZ vector tion containing an signal -factor sequence,. KEX2 cleavage a site (GVSLEKR) introduced was at the by junction. Charles H. Michael E. Streuli, Grant - 2000 Science - 360 pages - A pPICZ vector alpha1 reconstructed was with pPICZ alpha vector PCR and product of
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cerevisiae, for the targeting enzyme to the. by James M. - Cregg 2007 - Science - 268 pages Each pPICZ-derived containing vector or native mutant sequence for and GlcAT-P GlcAT-I
was transformed into the P. pastoris SMD1168 yeast strain using the. by Charles H. Streuli, Michael E. Grant - 2000