Journal: Jundishapur Journal of Microbiology

Article Title: Design, Construction and Cloning of Truncated ORF2 and tPA sp -PADRE-Truncated ORF2 Gene Cassette From Hepatitis E Virus in the pVAX1 Expression Vector

doi: 10.5812/jjm.26035

Figure Lengend Snippet: Schematic Model of the Recombinant tPA sp -PADRE-Truncated ORF2 Gene Cassette and Truncated ORF2 Gene

Article Snippet: Following verification of the construct’s properties, the optimized tPA sp -PADRE-truncated ORF2 gene cassette was then synthesized and cloned into the pBMH commercial cloning vector by Biomatik Company (Biomatik Corporation, Cambridge, Canada).

Techniques: Recombinant

Journal: Jundishapur Journal of Microbiology

Article Title: Design, Construction and Cloning of Truncated ORF2 and tPA sp -PADRE-Truncated ORF2 Gene Cassette From Hepatitis E Virus in the pVAX1 Expression Vector

doi: 10.5812/jjm.26035

Figure Lengend Snippet: A, Codon Adaptation Index of the non-optimized and optimized truncated ORF2 gene; B, GC content of the non optimized and optimized truncated ORF2 gene; C, Frequency of optimal codons (FOP) of the non optimized and optimized truncated ORF2 gene; D, Presence (left picture) and absence (right picture) of 22-amino acid secretory peptide before and after addition to truncated ORF2 protein using the signal peptide analysis online software.

Article Snippet: Following verification of the construct’s properties, the optimized tPA sp -PADRE-truncated ORF2 gene cassette was then synthesized and cloned into the pBMH commercial cloning vector by Biomatik Company (Biomatik Corporation, Cambridge, Canada).

Techniques: Software

Journal: Jundishapur Journal of Microbiology

Article Title: Design, Construction and Cloning of Truncated ORF2 and tPA sp -PADRE-Truncated ORF2 Gene Cassette From Hepatitis E Virus in the pVAX1 Expression Vector

doi: 10.5812/jjm.26035

Figure Lengend Snippet: A, Sub-cloning Results of Optimized tPA sp -PADRE-Truncated ORF2 (aa 112 - 660) Gene Cassette In Pvax1 Eukaryotic Plasmid; Lane M, 1kb DNA marker; Lane 1, pBMH-tPA sp -PADRE-truncated ORF2 plasmid; Lane 2, digested pBMH-tPA sp -PADRE-truncated ORF2 plasmid by NheI restriction enzyme; Lane 3, digested pBMH-tPA sp -PADRE-truncated ORF2 plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2900-bp and a 1795-bp fragment; Lane 4, pVAX plasmid; Lane 5, digested pVAX plasmid by NheI and XhoI restriction enzymes and production of 2909-bp band; Lane 6, pVAX-tPA sp -PADRE-truncated ORF2 plasmid; Lane 7, digested pVAX-tPA sp -PADRE-truncated ORF2 plasmid by the NheI restriction enzyme; Lane 8, digested pVAX-tPA sp -PADRE-truncated ORF2 plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2909-bp and a 1795-bp fragment; Lane 9, 1876-bp band in colony PCR test with T7 and BHG universal primers; B, Restriction Enzyme analyses and colony polymerase chain reaction results of three colonies containing pVAX-tPA sp -PADRE-truncated ORF2 plasmid; Lane M, 1 kb DNA marker; Lane 1, negative control; Lanes 2 - 4, 1876-bp band in colony PCR test; Lanes 5 - 7, digested pVAX-tPA sp -PADRE-truncated ORF2 (aa 112 - 660) -linker plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2909-bp and a 1795-bp fragment

Article Snippet: Following verification of the construct’s properties, the optimized tPA sp -PADRE-truncated ORF2 gene cassette was then synthesized and cloned into the pBMH commercial cloning vector by Biomatik Company (Biomatik Corporation, Cambridge, Canada).

Techniques: Subcloning, Plasmid Preparation, Marker, Polymerase Chain Reaction, Negative Control

Journal: Jundishapur Journal of Microbiology

Article Title: Design, Construction and Cloning of Truncated ORF2 and tPA sp -PADRE-Truncated ORF2 Gene Cassette From Hepatitis E Virus in the pVAX1 Expression Vector

doi: 10.5812/jjm.26035

Figure Lengend Snippet: A, Polymerase Chain Reaction Results of pVAX-Truncated ORF2 (aa 112 - 660) Expression Plasmid (without PADRE and tPA sp sequences); Lane M, 1 kb DNA marker; Lane 1, pVAX-truncated ORF2 (aa 112 - 660) plasmid; Lane 2, digested pVAX-truncated ORF2 (aa 112 - 660) plasmid by NheI restriction enzyme (4596 bp fragment); Lanes 3, digested pVAX-truncated ORF2 (112 - 660) plasmid by NheI and XhoI restriction enzymes and production of two expected fragments, a 2909-bp and a 1693-bp fragment; Lanes 4, PCR product of truncated ORF2 (aa 112-660) gene (without PADRE and tPA sp sequences) with specific primers (1710 bp); Lanes 4, PCR product of tPA sp -PADRE-ORF2 (aa 112 - 660) gene (with PADRE and tPA sp sequences) with T7p and BGH primers (1876 bp); B, Colony Polymerase Chain Reaction Results of Colonies Containing the pVAX-Truncated ORF2 (aa 112 - 660) Expression Recombinant Plasmid; Lane M, 1 kb DNA marker; Lane 1, negative control; Lanes 2 - 9, 1774 bp band of pVAX-truncated ORF2 (aa 112 - 660) plasmid (without PADRE and tPA sp sequences) due to colony PCR assay with T7p and BGH universal primers; Lane 2, 1876 bp band of pVAX-tPA sp -PADRE-ORF2 (aa 112 - 660) plasmid (with PADRE and tPA sp sequences) due to colony PCR assay with T7p and BGH primers as controls

Article Snippet: Following verification of the construct’s properties, the optimized tPA sp -PADRE-truncated ORF2 gene cassette was then synthesized and cloned into the pBMH commercial cloning vector by Biomatik Company (Biomatik Corporation, Cambridge, Canada).

Techniques: Polymerase Chain Reaction, Expressing, Plasmid Preparation, Marker, Recombinant, Negative Control

Journal: Jundishapur Journal of Microbiology

Article Title: Design, Construction and Cloning of Truncated ORF2 and tPA sp -PADRE-Truncated ORF2 Gene Cassette From Hepatitis E Virus in the pVAX1 Expression Vector

doi: 10.5812/jjm.26035

Figure Lengend Snippet: A,Reverse Transcription-Polymerase Chain Reaction Amplification of Truncated ORF2 (aa 112-660) Gene From Transfected Cells and Electrophoresis on 2% Agarose Gel; Lane M, 100 bp DNA marker; Lane 1, RT-PCR product of truncated ORF2 (112 - 660) gene by F1478 and R1730 specific primers with 271-bp length, from CHO cells transfected with expression recombinant plasmid pVAX-truncated ORF2 (aa 112 - 660) ; Lane 2, RT-PCR assay of truncated ORF2 (112 - 660) gene from CHO cells transfected with pVAX1 plasmid (negative control); Lane 3, RT-PCR assay of truncated ORF2 (112 - 660) gene from untransfected cells (negative control); B, Reverse transcription-polymerase chain reaction amplification of tPA sp -PADRE-Truncated ORF2 (aa 112 - 660) Gene From Transfected Cells and Electrophoresis on 2% Agarose Gel; Lane M, 100 bp DNA marker; Lane 1 and 6, RT-PCR assay of truncated ORF2 gene from untransfected cells (negative control); Lanes 2 and 3, RT-PCR product of truncated ORF2 (aa 112 - 660) gene by F1478 and R1730 specific primers with 271-bp length, from CHO and HEK293 cells transfected with expression recombinant plasmid pVAX-tPA sp -PADRE-truncated ORF2 (aa 112 - 660) ; Lanes 4 and 5, RT-PCR assay of truncated ORF2 gene from cells transfected with the pVAX plasmid (negative control).

Article Snippet: Following verification of the construct’s properties, the optimized tPA sp -PADRE-truncated ORF2 gene cassette was then synthesized and cloned into the pBMH commercial cloning vector by Biomatik Company (Biomatik Corporation, Cambridge, Canada).

Techniques: Reverse Transcription Polymerase Chain Reaction, Amplification, Transfection, Electrophoresis, Agarose Gel Electrophoresis, Marker, Expressing, Recombinant, Plasmid Preparation, Negative Control

Journal: Jundishapur Journal of Microbiology

Article Title: Design, Construction and Cloning of Truncated ORF2 and tPA sp -PADRE-Truncated ORF2 Gene Cassette From Hepatitis E Virus in the pVAX1 Expression Vector

doi: 10.5812/jjm.26035

Figure Lengend Snippet: A, Detection of HEV truncated ORF2 protein in CHO cells and B, HEK293 cells transfected with pVAX-tPA sp -PADRE-truncated ORF2 (aa 112 - 660) recombinant plasmid by the IFAT test. Detection of HEV ORF2 protein in CHO cells and C, HEK293 cells; D, transfected with pVAX-truncated ORF2 (aa 112 - 660) recombinant plasmid by the IFAT test.

Article Snippet: Following verification of the construct’s properties, the optimized tPA sp -PADRE-truncated ORF2 gene cassette was then synthesized and cloned into the pBMH commercial cloning vector by Biomatik Company (Biomatik Corporation, Cambridge, Canada).

Techniques: Transfection, Recombinant, Plasmid Preparation