m smegmatis mc 155 strain  (ATCC)

Journal: Archives of Microbiology

Article Title: Export of eDNA is a mechanism used by Mycobacterium avium subsp. hominissuis for survival within host macrophages

doi: 10.1007/s00203-025-04396-y

Figure Lengend Snippet: M. avium species suppresses NLRP3 expression and IL-1β secretion by THP-1 macrophages. Naive and LPS activated macrophages were infected with either M. avium 104, M. avium A5, or M. smegmatis to determine NLRP3 gene expression and IL-1β protein section over 48 h.p.i. NLRP3 gene expression in A naïve and C LPS activated THP-1macrophages quantified by qPCR. IL-1β protein levels in supernatants from B naive and D LPS activated THP-1 macrophages quantified by ELISA. Statistical comparisons represent significant differences from uninfected cells at each timepoint. Data are representative of three independent experiments. Statistical comparisons: *P < 0.05; **P < 0.005, ***P < 0.0005

Article Snippet: M. smegmatis mc 2 155 ( M. smegmatis ) was obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, Infection, Gene Expression, Enzyme-linked Immunosorbent Assay

Journal: Archives of Microbiology

Article Title: Export of eDNA is a mechanism used by Mycobacterium avium subsp. hominissuis for survival within host macrophages

doi: 10.1007/s00203-025-04396-y

Figure Lengend Snippet: M. avium species suppresses the expression of DNA sensors AIM2, cGAS, and STING, and triggers IFN-β secretion. Naïve and LPS activated macrophages were infected with either M. avium 104, M. avium A5, or M. smegmatis to determine DNA sensor gene expression and IFN-β protein production. AIM2 gene expression in naïve macrophages ( A ) and LPS activated macrophages ( B ) over 48 h.p.i. determined by qPCR. cGAS ( C ) and STING ( D ) gene expression after 24 h.p.i. quantified by qPCR. IFN-β protein levels ( E ) after 24 h.p.i. quantified by ELISA. Data are representative of at least two independent experiments. Statistical comparisons: *P < 0.05; **P < 0.005, ***P < 0.0005

Article Snippet: M. smegmatis mc 2 155 ( M. smegmatis ) was obtained from American Type Culture Collection (ATCC).

Techniques: Expressing, Infection, Gene Expression, Enzyme-linked Immunosorbent Assay

Journal: Applied and Environmental Microbiology

Article Title: Survival and spread of engineered Mycobacterium smegmatis and associated mycobacteriophage in soil microcosms

doi: 10.1128/aem.00212-25

Figure Lengend Snippet: M. smegmatis persistence at the inoculation site over 49 days. ( A ) M. smegmatis CFU/g dry soil was determined by colony counting in both sterilized (left) and non-sterilized (right) soil. M. smegmatis counts in control microcosms are shown in black, Ms- pC3 in blue, and Ms- pML(int) in red. Lines depict the average of 3 biological replicates, with each replicate displayed as an individual point. ( B ) M. smegmatis 16S rRNA relative abundance (as a percentage) in non-sterilized soil. M. smegmatis relative abundance in control microcosms is shown in black, Ms- pC3 in blue, and Ms- pML(int) in red. Lines depict the average of three biological replicates, with each replicate displayed as an individual point. ( C ) M. smegmatis shotgun metagenomic relative abundance (as a percentage) in non-sterilized soil on days 7 and 49. White circles show the mean of three biological replicates, with each replicate displayed as a blue ( Ms -pC3) or red ( Ms -pML(int)) point.

Article Snippet: Two different engineered strains of M. smegmatis mc 2 155 (ATCC #700084) were used in the microcosm experiments: one with mCherry, a red fluorescence protein gene, transformed in a plasmid (called Ms -pC3) and one with mCherry integrated into the genome (called Ms- pML(int)).

Techniques: Control

Journal: Applied and Environmental Microbiology

Article Title: Survival and spread of engineered Mycobacterium smegmatis and associated mycobacteriophage in soil microcosms

doi: 10.1128/aem.00212-25

Figure Lengend Snippet: M. smegmatis abundance at 5 cm away from the inoculation site. ( A ) M. smegmatis CFU/g dry soil at 5 cm from the inoculation site as measured by colony counting. M. smegmatis counts in control microcosms are shown in black, Ms- pC3 in blue, and Ms- pML(int) in red. Lines depict the average of three biological replicates, with each replicate displayed as an individual point. ( B ) M. smegmatis 16S rRNA relative abundances (as a percentage) in non-sterilized soil microcosms at 5 cm from the inoculation site. M. smegmatis relative abundances in control microcosms are shown in black, Ms- pC3 in blue, and Ms- pML(int) in red. Lines depict the average of three biological replicates, with each replicate displayed as an individual point.

Article Snippet: Two different engineered strains of M. smegmatis mc 2 155 (ATCC #700084) were used in the microcosm experiments: one with mCherry, a red fluorescence protein gene, transformed in a plasmid (called Ms -pC3) and one with mCherry integrated into the genome (called Ms- pML(int)).

Techniques: Control

Journal: Applied and Environmental Microbiology

Article Title: Survival and spread of engineered Mycobacterium smegmatis and associated mycobacteriophage in soil microcosms

doi: 10.1128/aem.00212-25

Figure Lengend Snippet: Kampy persistence at the inoculation site. ( A ) Kampy PFU/g dry soil at the inoculation site over 49 days. Kampy population density was measured by plaque counting on plates with M. smegmatis lawns. Kampy counts in control microcosms are shown in black, Kampy counts in Ms- pC3 microcosms in blue, and Kampy counts in Ms- pML(int) microcosms in red. Lines depict the average of 3 biological replicates, with each replicate displayed as an individual point. ( B ) The correlation between the relative abundance (as a percentage) of Kampy metagenomic shotgun sequencing reads and phage plaque counts at the inoculation site in non-sterilized soil on days 4 and 49. Soil microcosm replicates are represented by individual points, with the dotted line representing the linear correlation between the plaque assay measurement and the relative abundance of Kampy within each sample.

Article Snippet: Two different engineered strains of M. smegmatis mc 2 155 (ATCC #700084) were used in the microcosm experiments: one with mCherry, a red fluorescence protein gene, transformed in a plasmid (called Ms -pC3) and one with mCherry integrated into the genome (called Ms- pML(int)).

Techniques: Control, Shotgun Sequencing, Plaque Assay

Journal: Applied and Environmental Microbiology

Article Title: Survival and spread of engineered Mycobacterium smegmatis and associated mycobacteriophage in soil microcosms

doi: 10.1128/aem.00212-25

Figure Lengend Snippet: Kampy PFU/g dry soil 5 cm away from the inoculation site. Kampy density was measured by plaque counting on plates with M. smegmatis lawns. Kampy counts in control microcosms are shown in black, Kampy counts in Ms- pC3 microcosms in blue, and Kampy counts in Ms- pML(int) microcosms in red. Lines depict the average of 3 biological replicates, with each replicate displayed as an individual point.

Article Snippet: Two different engineered strains of M. smegmatis mc 2 155 (ATCC #700084) were used in the microcosm experiments: one with mCherry, a red fluorescence protein gene, transformed in a plasmid (called Ms -pC3) and one with mCherry integrated into the genome (called Ms- pML(int)).

Techniques: Control

Journal: Applied and Environmental Microbiology

Article Title: Survival and spread of engineered Mycobacterium smegmatis and associated mycobacteriophage in soil microcosms

doi: 10.1128/aem.00212-25

Figure Lengend Snippet: Microbial community structure after inoculation of engineered M. smegmatis and Kampy phage. Principal component analysis performed in R. ( A ) PCA on 16S rRNA sequencing reads from non-sterilized soil microcosms across all time points. ( B ) PCA on shotgun metagenomic sequencing reads from the inoculation site for non-sterilized soil microcosms on days 7 and 49.

Article Snippet: Two different engineered strains of M. smegmatis mc 2 155 (ATCC #700084) were used in the microcosm experiments: one with mCherry, a red fluorescence protein gene, transformed in a plasmid (called Ms -pC3) and one with mCherry integrated into the genome (called Ms- pML(int)).

Techniques: Sequencing

Journal: Analytical Chemistry

Article Title: Nano-Infrared Detection and Identification of Bacteria at the Single-Cell Level

doi: 10.1021/acs.analchem.5c01677

Figure Lengend Snippet: Nanoscale imaging and analysis of bacterial cells. (A) AFM images of B. burgdorferi , E. coli , M. smegmatis, and two strains of A. baumannii onto gold-coated silicon wafers. Scale bars are 5 μM ( B. burgdorferi ), 0.78 μM ( E. coli ), 3.11 μM ( A. baumannii ), and 1.71 μM ( M. smegmatis ). (B) Averaged AFM-IR spectra acquired from the bacteria.

Article Snippet: M. smegmatis strain mc 2 155 was purchased from ATCC (Lot no. 700084).

Techniques: Imaging, Bacteria

Journal: Analytical Chemistry

Article Title: Nano-Infrared Detection and Identification of Bacteria at the Single-Cell Level

doi: 10.1021/acs.analchem.5c01677

Figure Lengend Snippet: (A) Latent variable number vs CV and calculated (Cal) classification error average. (B) 3D LVs plot of acquired spectra of B. burgdorferi (pink), E. coli (blue), M. smegmatis (green), and two strains of A. baumannii (strain 1, red and strain 2, light blue); 95% confidence intervals are shown with colored ellipses.

Article Snippet: M. smegmatis strain mc 2 155 was purchased from ATCC (Lot no. 700084).

Techniques:

Journal: G3: Genes | Genomes | Genetics

Article Title: Genome-wide screen overexpressing mycobacteriophage Amelie genes identifies multiple inhibitors of mycobacterial growth

doi: 10.1093/g3journal/jkae285

Figure Lengend Snippet: Results of cytotoxicity assays for representative Amelie genes. a) Recombinant pExTra plasmids constructed in this study encode Amelie gene sequences downstream of the pTet promoter and upstream of mcherry. The 2 genes in this aTc-inducible operon are transcriptionally linked, with each gene having distinct start and stop codons and RBS for translation of the 2 protein products. b) Results of representative cytotoxicity assays are shown to demonstrate the range of observed growth defects. In each assay, colonies of Mycobacterium smegmatis mc 2 155 transformed with the specified pExTra plasmid were resuspended, serially diluted, and spotted on 7H10 Kan media containing 0, 10, or 100 ng/ml aTc. Triplicate colonies (A, B, C) were tested for each gene alongside a positive control strain (+) transformed with pExTra02 (expressing wild-type Fruitloop 52 ) and a negative control strain (−) transformed with pExTra03 (expressing Fruitloop 52 I70S ). Cytotoxicity score (0) is represented by Amelie 30 . Cytotoxicity score (1) is represented by Amelie 58. Cytotoxicity score (2) is represented by Amelie 50 . Cytotoxicity score (3) is represented by Amelie 47 .

Article Snippet: Amelie was propagated on M. smegmatis mc 2 155 grown at 37°C in the presence of 1 mM CaCl 2 and no Tween in Middlebrook media and top agar.

Techniques: Recombinant, Construct, Transformation Assay, Plasmid Preparation, Positive Control, Expressing, Negative Control

Journal: G3: Genes | Genomes | Genetics

Article Title: Genome-wide screen overexpressing mycobacteriophage Amelie genes identifies multiple inhibitors of mycobacterial growth

doi: 10.1093/g3journal/jkae285

Figure Lengend Snippet: The genome of phage Amelie. The Amelie genome is shown as a line with kbp markers and genes represented by boxes—those above the line are transcribed rightwards and those below are transcribed leftwards. Numbers inside the box correspond to gene numbers and predicted functions are indicated above or below each gene. Box shading corresponds to cytotoxicity scoring, with white boxes designating genes found to have no effect on M. smegmatis growth (cytotoxicity score 0 ), the gray box indicating that clones for the gene were not recovered (NC) and thus was not tested in this study, the checkered box indicating a gene for which transformants (NT) could not be recovered and purple representing observed toxicity in our assay. The saturation of purple boxes corresponds to the severity of growth inhibition using the following scores: light purple (score 1 ; reduction in colony size; genes 8 , 16 , 46 , 55 , 56 , 58 , 63 , 65 , and 74 ), medium purple (score 2 ; 1–3 log reduction in viability; genes 9 , 15 , 20 , 31 , 32 , 50 , and 62 ), and dark purple (score 3 ; > 3-log reduction in viability; genes 17 , 29 , 34 , 43 , 44 , 47 , 49 , 57 , and 73 ).

Article Snippet: Amelie was propagated on M. smegmatis mc 2 155 grown at 37°C in the presence of 1 mM CaCl 2 and no Tween in Middlebrook media and top agar.

Techniques: Clone Assay, Inhibition

Journal: G3: Genes | Genomes | Genetics

Article Title: Genome-wide screen overexpressing mycobacteriophage Amelie genes identifies multiple inhibitors of mycobacterial growth

doi: 10.1093/g3journal/jkae285

Figure Lengend Snippet: Amelie genes observed to inhibit Mycobacterium smegmatis growth upon overexpression.

Article Snippet: Amelie was propagated on M. smegmatis mc 2 155 grown at 37°C in the presence of 1 mM CaCl 2 and no Tween in Middlebrook media and top agar.

Techniques: Over Expression, Scaffolding

Journal: JACS Au

Article Title: A Systematic Approach to Discover New Natural Product Scaffolds Using Database-Derived Relative Mass Spectral Defects and Molecular Networking

doi: 10.1021/jacsau.4c00889

Figure Lengend Snippet: MICs of Brasiliencins A ( 1 ) and B ( 2 ) against Mycobacterium smegmatis MC 2 155

Article Snippet: The major compound brasiliencin A was extremely potent in inhibiting the growth of M. smegmatis MC 155 (ATCC 700084) with an MIC of 31.3 nM and inhibited the growth of the oral microbiome-derived Streptococcus australis at low micromolar concentrations (MIC of 7.8 μM).

Techniques: