ppiczαa expression construct plasmid dna  (Thermo Fisher)

Journal: Animals : an open access journal from MDPI

Article Title: Porcine β-Defensin 2 Expressed in Pichia pastoris Alleviates Enterotoxigenic Escherichia coli -Induced Intestinal Injury and Inflammatory Response in Mice.

doi: 10.3390/ani15101389

Figure Lengend Snippet: Figure 1. Construction of the PBD2 expression plasmid and its P. pastoris transformants. (A) Codon- optimized cDNA sequence of PBD2 for P. pastoris (upper line) and its corresponding amino acid sequence (lower line). *, stop codon. (B) Schematic diagram of recombinant plasmid pPPBD2. (C) PCR identification of pPPBD2. M, Marker; 1–2, plasmid pPICZαA; 3–4, pPPBD2; 5, negative control. (D) Identification of P. pastoris transformants that have the highest inhibition rate against ATCC 29213. M, Marker; 1–2, positive transformants; 3–4, P. pastoris X33; 5–6, pPPBD2 plasmid; 7, negative control.

Article Snippet: M, Marker; 1–2, plasmid pPICZαA; 3–4, pPPBD2; 5, negative control. (D) Identification of P. pastoris transformants that have the highest inhibition rate against ATCC 29213.

Techniques: Expressing, Plasmid Preparation, Sequencing, Recombinant, Marker, Negative Control, Inhibition

Journal: Animals : an open access journal from MDPI

Article Title: Porcine β-Defensin 2 Expressed in Pichia pastoris Alleviates Enterotoxigenic Escherichia coli -Induced Intestinal Injury and Inflammatory Response in Mice.

doi: 10.3390/ani15101389

Figure Lengend Snippet: Figure 2. Expression of PBD2 in P. pastoris. (A) Preliminary screening of positive transformants. In total, 100 µL supernatant of different transformants was co-incubated with 100 µL S. aureus ATCC 29213 with an OD600 of 0.02, and the OD600 was measured after 8 h. Inhibition ratios greater than 15% are shown. (B) Secondary screening of positive transformants. In total, 500 µL supernatant was co-incubated with 500 µL ATCC 29213 (5 × 105 CFU/mL) for 1 h at 37 ◦C without shaking, and 100 µL from each sample was withdrawn to count the bacterial number. (C) Tricine SDS- PAGE analysis of rPBD2-51 with 1% methanol induction for 72 h. M, Marker; 1, 15 µL supernatant from the transformant which harbors the pPICZαA vector; 2, 15 µL supernatant from rPBD2-51. (D) Western blot analysis of rPBD2-51 using PBD2 monoclonal antibody. M, Marker; 1, pPICZαA vector supernatant; 2, rPBD2-51 supernatant. (E) The rPBD2 supernatant inhibits the growth of ATCC 29213. 1, The supernatant from the transformant which harbors the pPICZαA vector; 2, The supernatant from rPBD2-51; 3, Ampicillin.

Article Snippet: M, Marker; 1–2, plasmid pPICZαA; 3–4, pPPBD2; 5, negative control. (D) Identification of P. pastoris transformants that have the highest inhibition rate against ATCC 29213.

Techniques: Expressing, Incubation, Inhibition, SDS Page, Marker, Plasmid Preparation, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Heterologous Production, Purification and Characterization of Two Cold-Active β- d -Galactosidases with Transglycosylation Activity from the Psychrotolerant Arctic Bacterium Arthrobacter sp. S3* Isolated from Spitsbergen Island Soil

doi: 10.3390/ijms252413354

Figure Lengend Snippet: Summary of the purification of recombinant Arthrobacter sp. S3* Bgal2 and Bgal42 β- d -galactosidases obtained from 1 L of E. coli LMG194 culture.

Article Snippet: The resulting pPICZαA/GH42βGalS3*I recombinant plasmid was linearized with Mss I ( Pme I) enzyme and transformed into K. phaffii X-33 cells (Invitrogen, Carlsbad, CA, USA) by electroporation.

Techniques: Purification, Recombinant, Activity Assay