Journal: Journal of Virology

Article Title: Sulfolobus Turreted Icosahedral Virus c92 Protein Responsible for the Formation of Pyramid-Like Cellular Lysis Structures ^▿ †

doi: 10.1128/JVI.00379-11

Figure Lengend Snippet: Schematics. (A) c92 gene cloned into pSeSD1 overexpression vector showing the location of the FLAG and His tags on the C-terminal end of the protein (underlined in the amino acid sequence); the NdeI (CATATG) and SalI (GTCGAC) sites are underlined in the nucleotide sequence. (B) The c92 c53a mutant construct, showing the change of a serine residue (TCA) to a premature stop codon (TAA); the transmembrane domain is underlined in the amino acid sequence.

Article Snippet: The PCR conditions were as follows: 5× GoTaq buffer (Promega), 10 mM dNTPs, 50 pmol/μl c92-SEQ-F primer (5′-ACCCGAGAATGAGGTGAAGC-3′), 50 pmol/μl c92-SEQ-R primer (5′-CAGTAGGTACCCGGGAAAAA-3′), 1 U GoTaq DNA polymerase (Promega), and 2 μl of the transformed Sulfolobus PH1-16 culture.

Techniques: Clone Assay, Over Expression, Plasmid Preparation, Sequencing, Mutagenesis, Construct, Residue

Journal: Journal of Virology

Article Title: Sulfolobus Turreted Icosahedral Virus c92 Protein Responsible for the Formation of Pyramid-Like Cellular Lysis Structures ^▿ †

doi: 10.1128/JVI.00379-11

Figure Lengend Snippet: (A) Western blot of PH1-16 cultures with the c92-FLAG construct. M represents protein size standard; lane 1, preinduction; lane 2, arabinose induced (t = 44 h); lane 3, galactose repressed (t = 44 h). (B) Western blot showing the localization of the c92 protein in PH1-16 cells. M represents protein size standard; lane 1, membrane fraction; lane 2, insoluble fraction; lane 3, soluble fraction.

Article Snippet: The PCR conditions were as follows: 5× GoTaq buffer (Promega), 10 mM dNTPs, 50 pmol/μl c92-SEQ-F primer (5′-ACCCGAGAATGAGGTGAAGC-3′), 50 pmol/μl c92-SEQ-R primer (5′-CAGTAGGTACCCGGGAAAAA-3′), 1 U GoTaq DNA polymerase (Promega), and 2 μl of the transformed Sulfolobus PH1-16 culture.

Techniques: Western Blot, Construct, Membrane

Journal: Journal of Virology

Article Title: Sulfolobus Turreted Icosahedral Virus c92 Protein Responsible for the Formation of Pyramid-Like Cellular Lysis Structures ^▿ †

doi: 10.1128/JVI.00379-11

Figure Lengend Snippet: (A) Arabinose-induced Sulfolobus PH1-16 cultures overexpressing c92 protein harvested at 44 h after induction (arrows point to pyramid structures). (B) Galactose-repressed Sulfolobus PH1-16 cells harvested at 44 h after repression. (C) Immunolocalization of overexpressed c92 in Sulfolobus PH1-16 showing localization to pyramid-like lysis structures (red arrows indicate gold particles on pyramid structure, and black arrows indicate internal membrane structures near the base of the pyramids). (D) Immunolocalization of overexpressed c92 in Sulfolobus PH1-16 showing localization to internal membranes (black arrows indicate internal membrane structure and red arrows indicate gold particles). (E) Galactose-repressed Sulfolobus cells showing no gold particles. Bars, 250 nm (A and B) or 100 nm (C to E).

Article Snippet: The PCR conditions were as follows: 5× GoTaq buffer (Promega), 10 mM dNTPs, 50 pmol/μl c92-SEQ-F primer (5′-ACCCGAGAATGAGGTGAAGC-3′), 50 pmol/μl c92-SEQ-R primer (5′-CAGTAGGTACCCGGGAAAAA-3′), 1 U GoTaq DNA polymerase (Promega), and 2 μl of the transformed Sulfolobus PH1-16 culture.

Techniques: Lysis, Membrane

Journal: Journal of Virology

Article Title: Sulfolobus Turreted Icosahedral Virus c92 Protein Responsible for the Formation of Pyramid-Like Cellular Lysis Structures ^▿ †

doi: 10.1128/JVI.00379-11

Figure Lengend Snippet: Model of c92 function in the replication cycle of STIV. (A) Schematic of early viral replication indicating DNA replication, transcription, and translation of viral proteins. (B) The STIV c92 protein results in the formation of pyramid-like lysis structures and internal membrane vesicles. (C) Internal membrane structures serve as templates for STIV assembly. (D) Cell lyses to release viral progeny.

Article Snippet: The PCR conditions were as follows: 5× GoTaq buffer (Promega), 10 mM dNTPs, 50 pmol/μl c92-SEQ-F primer (5′-ACCCGAGAATGAGGTGAAGC-3′), 50 pmol/μl c92-SEQ-R primer (5′-CAGTAGGTACCCGGGAAAAA-3′), 1 U GoTaq DNA polymerase (Promega), and 2 μl of the transformed Sulfolobus PH1-16 culture.

Techniques: Lysis, Membrane