Journal: Viruses

Article Title: Biological Activity of Optimized Codon Bovine Type III Interferon Expressed in Pichia pastoris

doi: 10.3390/v15051101

Figure Lengend Snippet: Construction of pPICZαA–boIFN–λ3 by SOE with optimized codons.

Article Snippet: The host strain GS115 and the expression vector pPICZαA were obtained from Invitrogen (Carlsbad, CA, USA).

Techniques:

Journal: Viruses

Article Title: Biological Activity of Optimized Codon Bovine Type III Interferon Expressed in Pichia pastoris

doi: 10.3390/v15051101

Figure Lengend Snippet: Identification of boIFN–λ3/λ3 V18M . ( A , B ). boIFN–λ3 and boIFN–λ3 V18M SOE–PCR, respectively. M: Trans2K DNA Marker; 1~4, 5~6, 7, and 8 indicate the first, second, third, and fourth PCR results, respectively. ( C ). Enzyme digestion and PCR identification of the P. pastoris transformants integrated into the boIFN–λ3 gene. M. Trans2K PlusII DNA Marker; 1 and 5. PCR result based on P1/P14; 2 and 6. Xho I digested; 3 and 7 Xho I and Xba I digested; 4 and 8. PCR result based on 5′AOX/P14. 1~4 was the result of recombinant expression vector pPICZαA–boIFN–λ3 and 5~8 indicated the pPICZαA–boIFN–λ3 V18M .

Article Snippet: The host strain GS115 and the expression vector pPICZαA were obtained from Invitrogen (Carlsbad, CA, USA).

Techniques: Overlap Extension Polymerase Chain Reaction, Marker, Recombinant, Expressing, Plasmid Preparation

Journal: Viruses

Article Title: Biological Activity of Optimized Codon Bovine Type III Interferon Expressed in Pichia pastoris

doi: 10.3390/v15051101

Figure Lengend Snippet: Selection of highly expressed recombinant GS115–pPICZαA–boIFN–λ3. The numbers 1, 2, 3, 5, 6, and 13 indicate recombinant GS115–pPICZαA–boIFN–λ3, and NC indicates GS115–pPICZαA. ( A ). Identification results of expressed recombinant GS115–pPICZαA–boIFN–λ3 by Western blot. ( B ). The results of supernatants of pPICZαA–boIFN–λ3 were identified by ELISA.

Article Snippet: The host strain GS115 and the expression vector pPICZαA were obtained from Invitrogen (Carlsbad, CA, USA).

Techniques: Selection, Recombinant, Western Blot, Enzyme-linked Immunosorbent Assay

Journal: Viruses

Article Title: Biological Activity of Optimized Codon Bovine Type III Interferon Expressed in Pichia pastoris

doi: 10.3390/v15051101

Figure Lengend Snippet: Optimization of the expression conditions of the recombinant boIFN–λ3/λ3 V18M . ( A , C ) The SDS–PAGE analysis of different induction times of the recombinant boIFN–λ3 and boIFN–λ3 V18M , respectively. ( B , D ). The protein weight analysis of different induction times of recombinant boIFN–λ3 and boIFN–λ3 V18M , respectively. ( E , F ). SDS–PAGE analysis of recombinant P. pastoris GS115 under different methanol concentrations. ( E ). GS115–pPICZαA–boIFN–λ3. ( F ). GS115–pPICZαA–boIFN–λ3 V18M . M. Unstained protein molecular weight Marker; NC. The induced protein of GS115–pPICZαA.

Article Snippet: The host strain GS115 and the expression vector pPICZαA were obtained from Invitrogen (Carlsbad, CA, USA).

Techniques: Expressing, Recombinant, SDS Page, Molecular Weight, Marker

Journal: Viruses

Article Title: Biological Activity of Optimized Codon Bovine Type III Interferon Expressed in Pichia pastoris

doi: 10.3390/v15051101

Figure Lengend Snippet: Protein identification and glycoprotein analysis of recombinant boIFN–λ3/λ3 V18M . ( A ). SDS–PAGE for recombinant boIFN–λ3 strains 5 and 13. M. Unstained protein molecular weight Marker, 1. The induced protein of GS115–pPICZαA, 2 and 3 is the induced protein of GS115–pPICZαA–boIFN–λ3 strains 5 and 13, respectively. ( B , C ). Western blot analysis of rabbit anti bovine IFN–λ3 polyclonal antibody ( B ) and rabbit anti human IFN–λ3 polyclonal antibody ( C ) for immune response of recombinant boIFN–λ3. M. PageRuler Marker; 1. The induced protein of GS115–pPICZαA; 2,3. The induced protein of GS115–pPICZαA–boIFN–λ3 strains 5 and 13, respectively. ( D ). SDS–PAGE for recombinant boIFN–λ3 V18M strain 2. M. Unstained protein molecular weight Marker; 1. The induced protein of GS115–pPICZαA–boIFN–λ3 V18M strain 2. ( E , F ). Western blot analysis of rabbit anti bovine IFN–λ3 polyclonal antibody ( E ) and rabbit anti human IFN–λ3 polyclonal antibody ( F ) for immune response of recombinant boIFN–λ3 V18M . M. PageRuler Marker; 1. The induced protein of GS115–pPICZαA; 2. The induced protein of GS115–pPICZαA–boIFN–λ3 V18M strain 2. ( G ). SDS–PAGE analysis of the purification of recombinant boIFN–λ3 and boIFN–λ3 V18M . ( H ). Glycoprotein staining of recombinant boIFN–λ3 and boIFN–λ3 V18M .

Article Snippet: The host strain GS115 and the expression vector pPICZαA were obtained from Invitrogen (Carlsbad, CA, USA).

Techniques: Recombinant, SDS Page, Molecular Weight, Marker, Western Blot, Purification, Staining