Journal: Journal of the American Chemical Society

Article Title: Biosynthesis of 14-membered cyclopeptide alkaloids via non-heme-iron- and 2-oxoglutarate-dependent oxidative decarboxylation

doi: 10.1021/jacs.5c23400

Figure Lengend Snippet: ( A ) EIC traces of 3 and decarboxylated cyFPIY ( 4 ) in methanolic extracts of transgenic N. benthamiana leaves expressing ZjuPrec-(FPIY) 3 -ZjuBURP with or without candidate decarboxylase ZjuDC for 7 days. ( B ) In vitro reconstitution of ZjuDC activity in the presence of Fe(II) and 2OG. ( C ) In vitro cofactor and substrate screening assays of ZjuDC. ( D ) ZjuDC reaction. ( E ) Kinetic characterization of ZjuDC with substrate 3 . Abbreviations: 2OG - 2-oxoglutarate, EIC – extracted ion chromatogram. ( F ) ZjuDC reaction with dimethylated substrate N,N -Me 2 -cyFPIY ( 6 ).

Article Snippet: Assays were 50 μL total volume with final concentrations of 50 mM HEPES pH 7.5, 200 μM FeSO 4 -7H 2 O, 500 μM 2-oxoglutarate (Chem-Impex Int’l Inc.), 400 μM L-ascorbic acid sodium salt (Acros Organics), 4% dimethyl sulfoxide, and 90 μM substrate 3 .

Techniques: Transgenic Assay, Expressing, In Vitro, Activity Assay

Journal: Heliyon

Article Title: Persistent viruses in mosquito cultured cell line suppress multiplication of flaviviruses

doi: 10.1016/j.heliyon.2018.e00736

Figure Lengend Snippet: Identification of viruses infected in mosquito cultured cell line C6/36. A : Schematics of Shinobi tetravirus (SHTV) and Menghai rhabdovirus (MERV) genomic sequences. Encoded putative genes and their expected sizes are presented (RdRp: RNA-dependent RNA polymerase, VPg: Viral protein genome-linked, CP: capsid protein, N: nucleoprotein, P: phosphoprotein, M: matrix protein, G: glycoprotein, and L: RNA-dependent RNA polymerase). B : Phylogenetic analysis of SHTV and other viruses containing Permutotetraviridae . The amino acid sequences of putative RdRp were aligned and then analyzed using a maximum-likelihood method. Bootstrap replications are shown on the branches. C : Phylogenetic analysis of MERV and other rhabdoviruses. The amino acid sequences of L proteins were analyzed as described. D : Detection of SHTV or MERV in C6/36 JCRB strain and ECACC strain. The viral genome was determined by RT-PCR with the template RNA extracted from cell culture supernatant or cultured cells. PCR amplification from genomic DNA extracted from cultured cells were also determined. J: C6/36 JCRB strain, E: C6/36 ECACC strain. The original image is shown in Supplementary Material 1.

Article Snippet: The growth kinetics of flaviviruses in C6/36 JCRB strain was significantly lower than that in C6/36 ECACC strain ( A).

Techniques: Infection, Cell Culture, Genomic Sequencing, Reverse Transcription Polymerase Chain Reaction, Amplification

Journal: Heliyon

Article Title: Persistent viruses in mosquito cultured cell line suppress multiplication of flaviviruses

doi: 10.1016/j.heliyon.2018.e00736

Figure Lengend Snippet: C6/36 cells of JCRB and ECACC strains showed different susceptibility to flavivirus infection. Growth kinetics of arboviruses in C6/36 cell lines. A : C6/36 cells of either JCRB or ECACC strain were infected with Zika virus (ZIKV), Japanese encephalitis virus (JEV), or Dengue virus (DENV) at a multiplicity of infection (MOI) of 0.01, and the titers in the supernatants were determined by plaque assay. B : C6/36 cells of either JCRB or ECACC strain were infected with Getah virus (GETV) or Sindbis virus (SINV) with MOI = 0.001. Red lines show the titer in C6/36 JCRB strain and black lines show that in C6/36 ECACC strain. The experiment was triplicated, and the standard deviations are also indicated. Asterisks showed the results of t -test for the data group at each time points (* p < 0.05, ** p < 0.01).

Article Snippet: The growth kinetics of flaviviruses in C6/36 JCRB strain was significantly lower than that in C6/36 ECACC strain ( A).

Techniques: Infection, Virus, Plaque Assay