P. pastoris strain GS115 and plasmid pPIC9 were purchased from Invitrogen (USA, Catalog number: K171001). Escherichia coli Top10 competent cells were procured from Tiangen (Beijing, China). HisTrap™ FF prepack columns (5 mL) were GE products (USA). Frozen-EZ Yeast Transformation II Kit™ was from Zymo Reasearch (USA). Mouse IL-2 and IFN-gamma ELISA kits were bought from Beyotime (Shanghai, China). Restriction endonucleases, T4 DNA ligase and agarose gel DNA purification kits were Takara (Japan) products. Concanavalin A (ConA) and biotin were from Sigma (USA). Thiazolyl blue and yeast nitrogen base with ammonium sulfate without amino acids (YNB) were purchased from Genview (USA). All the other chemicals were of analytical grade. Gene synthesis and sequencing were carried out by Sangon (Shanghai, China).
Plasmid ppic9
Manufactured by Thermo Fisher Scientific
Sourced in United States
Plasmid pPIC9 is a vector used for the expression of recombinant proteins in the methylotrophic yeast Pichia pastoris. It contains the AOX1 promoter for methanol-inducible expression, the HIS4 gene for selection in Pichia, and the pUC19 bacterial origin of replication and ampicillin resistance gene for propagation in E. coli.
Automatically generated - may contain errors
Lab products found in correlation
Restriction endonuclease, by Takara Bio (2 mentions)
Escherichia coli trans1 t1, by Transgene (2 mentions)
Soluble starch, by Merck Group (2 mentions)
P pastoris gs115, by Thermo Fisher Scientific (2 mentions)
T4 dna ligase, by Takara Bio (1 mentions)
Agarose gel dna purification kit, by Takara Bio (1 mentions)
Concanavalin a (cona), by Merck Group (1 mentions)
Escherichia coli top10 competent cells, by Tiangen Biotech (1 mentions)
Thiazolyl blue, by Genview (1 mentions)
Restriction endonuclease, by New England Biolabs (1 mentions)
Plasmid extraction kit, by Tiangen Biotech (1 mentions)
Dna purification kit, by Takara Bio (1 mentions)
Dna isolation kit, by Tiangen Biotech (1 mentions)
La taq dna polymerase, by Takara Bio (1 mentions)
Fast mutagenesis system, by Transgene (1 mentions)
Fastpfu fly dna polymerase, by Transgene (1 mentions)
T4 dna ligase, by New England Biolabs (1 mentions)
Endoglycosidase h endo h, by New England Biolabs (1 mentions)
Glycogen, by Merck Group (1 mentions)
Genome walking kit, by Takara Bio (1 mentions)
5 protocols using plasmid ppic9
1
Recombinant Protein Production in Pichia pastoris
2
Heterologous Expression of Bacterial Enzymes
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Escherichia coli TransI-T1 was obtained from TransGen (Beijing, China) and used as the host for plasmid construction. Pichia pastoris GS115 and plasmid pPIC9 from Invitrogen (Carlsbad, CA, USA) were used for gene expression. Plasmid extraction kits were purchased from TIANGEN (Beijing, China), and restriction endonucleases were purchased from New England Biolabs (Ipswich, MA, USA). The DNA isolation kit and DNA polymerase pfu were purchased from TIANGEN. The DNA purification kit and LA Taq DNA polymerase were purchased from TaKaRa (Tsu, Japan). FastPfu Fly DNA polymerase, assembly kit, and fast mutagenesis system were purchased from TransGen. All other chemicals were of analytical grade and commercially available. Substrates such as soluble starch were purchased from Sigma-Aldrich (St. Louis, MO, USA).
3
Brucella Protein Expression in Yeast
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B. melitensis 16M strain and B. melitensis M5 vaccine were purchased from the China Institute of Veterinary Drug Control (Beijing, China). P. pastoris GS115 and plasmid pPIC9 were purchased from Invitrogen (Carlsbad, CA, USA). E. coli DH5α and pET-28a (+) vector were preserved in our laboratory. The secondary antibodies were purchased from Abcam (Shanghai, China). All media were prepared according to Pichia and E. coli expressions manuals. The protection experiment was conducted in a biosafety level 3 (BSL-3) laboratory.
4
Recombinant IBDV VP2 Subunit Vaccine
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The vv IBDV GX8/99 strain preserved in our laboratory was isolated from Guangxi and has been adapted in chicken embryo fibroblast cells. P. pastoris GS115 and plasmid pPIC9 were purchased from Invitrogen (Carlsbad, CA, USA). Commercial VP2 subunit vaccine was acquired from Qingdao Yebio Biological Engineering Co. Ltd. All media were prepared according to the manuals of Pichia expression.
5
Glucoamylase production in Bispora sp.
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Bispora sp. MEY-1 (CGMCC 2500; the China General Microbiological Culture Collection Center, Beijing, China) was cultivated in wheat bran medium [19] (link) at 30°C for 3–5 days to induce the production of glucoamylase as described previously [22] (link). Escherichia coli Trans1-T1 and the plasmid pEASY-T3 from TransGen (Beijing, China) were used for gene cloning and sequencing. Pichia pastoris GS115 and plasmid pPIC9 (Invitrogen, Carlsbad, CA, USA) were used for gene expression. P. pastoris expression media were prepared as described in the Pichia expression kit manual (Invitrogen).
Maltooligosaccharides, 4-nitrophenyl-α-d -maltopentaoside (p-NPG5), amylose, amylopectin, glycogen, soluble starch, dextran and isomaltose were purchased from Sigma-Aldrich (St. Louis, MO, USA). The Genome walking kit and restriction endonucleases were from TaKaRa (Otsu, Japan). T4 DNA ligase and endoglycosidase H (Endo H) were from New England Biolabs (Hitchin, UK). All other chemicals were of analytical grade and commercially available.
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Bispora sp. MEY-1 (CGMCC 2500; the China General Microbiological Culture Collection Center, Beijing, China) was cultivated in wheat bran medium [19] (link) at 30°C for 3–5 days to induce the production of glucoamylase as described previously [22] (link). Escherichia coli Trans1-T1 and the plasmid pEASY-T3 from TransGen (Beijing, China) were used for gene cloning and sequencing. Pichia pastoris GS115 and plasmid pPIC9 (Invitrogen, Carlsbad, CA, USA) were used for gene expression. P. pastoris expression media were prepared as described in the Pichia expression kit manual (Invitrogen).
Maltooligosaccharides, 4-nitrophenyl-α-
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