Journal: mLife

Article Title: Artificial intelligence‐assisted automatic and index‐based microbial single‐cell sorting system for One‐Cell‐One‐Tube

doi: 10.1002/mlf2.12047

Figure Lengend Snippet: AI‐assisted single‐cell sorting and proliferation. (A) Confusion matrix of the model in recognizing the cells. Background FP is misidentifying the background as the target and Background FN is misidentifying the target as the background. (B) Automatic identification of the target yeast cells. Scale bar = 30 μm. (C) Survival rate calculated by single‐cell culture and the sorting accuracy calculated based on single‐cell genome sequences. AI, artificial intelligence; FN, false negative; FP, false positive; NJ, noncarotenoid‐producing Saccharomyces cerevisiae BY4742; SS, carotenoid‐producing Phaffia rhodozyma ATCC 24202.

Article Snippet: Budding yeast strains S. cerevisiae BY4742 (designated as NJ) and P. rhodozyma ATCC 24202 (designated as SS) were cultured on yeast extract‐peptone‐dextrose (YPD) agar plates at 30°C for 24 h. A single colony was inoculated into PBS solution and diluted to a concentration of ~10 7 cell/ml.

Techniques: FACS, Cell Culture

Journal: mBio

Article Title: Identification of Multiple Cryptococcal Fungicidal Drug Targets by Combined Gene Dosing and Drug Affinity Responsive Target Stability Screening

doi: 10.1128/mBio.01073-16

Figure Lengend Snippet: Screening results for yeast colonies resistant to bithionol. (A) Bithionol fungicidal activity against Cryptococcus neoformans (H99) or Saccharomyces cerevisiae (BY4741). Two hundred microliters of a suspension of 1,000 cells/ml in asparagine and bithionol was added to microcentrifuge tubes. Cells were then incubated at 37°C with shaking for 24 h. Samples from each tube were plated on YPD medium and incubated in a 37°C incubator for 48 h, and colonies were counted. Results are from 3 independent assays; error bars represent standard deviations. Shown on the right is the chemical structure of bithionol. (B) Target identification for potential bithionol-interacting proteins by DARTS. S. cerevisiae lysates were incubated with or without bithionol for 1 h. Each sample was subjected to proteolysis. The partially digested proteins were separated on a 10% bis-Tris gel and visualized with silver staining. Two specific protein bands (indicated by arrows) were further processed for mass spectrum analysis. (C) Growth on plates was compared using replica plating of a 9,000-cell mixture cultured on agar with or without 10 µM bithionol at 37°C for 2 days. (D) Venn diagram of potential target genes from the gene dosing (GD) and drug affinity responsive target stability (DARTS) screen.

Article Snippet: Saccharomyces cerevisiae strain BY4741 and Cryptococcus neoformans strain H99 (ATCC 208821; a kind gift of J.

Techniques: Activity Assay, Suspension, Incubation, Drug discovery, Silver Staining, Cell Culture

Journal: mBio

Article Title: Identification of Multiple Cryptococcal Fungicidal Drug Targets by Combined Gene Dosing and Drug Affinity Responsive Target Stability Screening

doi: 10.1128/mBio.01073-16

Figure Lengend Snippet: Fungicidal assay results for S. cerevisiae overexpression strains exposed to increasing concentrations of bithionol. The indicated overexpressed strains and the WT BY4741 strain were used, and assays to determine CFU were performed in triplicate. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001.

Article Snippet: Saccharomyces cerevisiae strain BY4741 and Cryptococcus neoformans strain H99 (ATCC 208821; a kind gift of J.

Techniques: Over Expression

Journal: Indian Journal of Microbiology

Article Title: Homologous Recombination is Activated at Early Time Points Following Exposure to Cobalt Chloride Induced Hypoxic Conditions in Saccharomyces cerevisiae

doi: 10.1007/s12088-011-0195-1

Figure Lengend Snippet: Yeast strains used in this study

Article Snippet: At a concentration of 0.75 mM, CoCl 2 induced hypoxia-specific Ole1p expression without having any effect on cell viability [ 17 ]. table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Strains Genotype Source BY4741 Mat a his3 Δ leu2 Δ 0 met15 Δ 0 ura3 Δ 0 Dr. A. Bachhawat IMTECH, Chandigarh India D7 Mat a ade2 - 40 ilv1 - 92 trp5 - 12 Dr. Madhubala INMAS, Delhi, India Mat α ade2 - 119 ilv1 - 92 trp5 - 27 ATCC 201388 MAT a his3 Δ 1leu2 Δ 0 met15 Δ 0 ura3 Δ 0 Invitrogen RAD52 - GFP RAD52 - GFP - HIS3MX6 Open in a separate window Yeast strains used in this study Rate of Mitotic Recombination Rates of mitotic recombination were determined by fluctuation analysis.

Techniques:

Journal: Indian Journal of Microbiology

Article Title: Homologous Recombination is Activated at Early Time Points Following Exposure to Cobalt Chloride Induced Hypoxic Conditions in Saccharomyces cerevisiae

doi: 10.1007/s12088-011-0195-1

Figure Lengend Snippet: CoCl2 induces DNA damage and inhibits DNA replication. (A) CoCl2 induced DNA damage: Log phase BY4741 cells were either exposed to 0.1 mM H2O2, 0.1% MMS and 0.75 mM CoCl2 for 30 min or left unexposed (control) before proceeding for the comet assay. H2O2 and MMS were used as positive controls for the experiment. 50 individual comets were examined for each condition and the distribution of the Olive tail moment for each exposure condition is shown. (B) Same as (A) except that the data has been plotted in logarithmic scale to depict the low values of olive moment. (C) Cell cycle progression in the presence and absence of CoCl2: The experiment was performed thrice and representative data from a single experiment is shown

Article Snippet: At a concentration of 0.75 mM, CoCl 2 induced hypoxia-specific Ole1p expression without having any effect on cell viability [ 17 ]. table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Strains Genotype Source BY4741 Mat a his3 Δ leu2 Δ 0 met15 Δ 0 ura3 Δ 0 Dr. A. Bachhawat IMTECH, Chandigarh India D7 Mat a ade2 - 40 ilv1 - 92 trp5 - 12 Dr. Madhubala INMAS, Delhi, India Mat α ade2 - 119 ilv1 - 92 trp5 - 27 ATCC 201388 MAT a his3 Δ 1leu2 Δ 0 met15 Δ 0 ura3 Δ 0 Invitrogen RAD52 - GFP RAD52 - GFP - HIS3MX6 Open in a separate window Yeast strains used in this study Rate of Mitotic Recombination Rates of mitotic recombination were determined by fluctuation analysis.

Techniques: Control, Single Cell Gel Electrophoresis