Journal: International Journal of Molecular Sciences

Article Title: Whole Genome Sequencing and CRISPR/Cas9 Gene Editing of Enterotoxigenic Escherichia coli BE311 for Fluorescence Labeling and Enterotoxin Analyses

doi: 10.3390/ijms23147502

Figure Lengend Snippet: The GFP-expressing ability of different E. coli strains and vectors.

Article Snippet: The BE311 strain was deposited in the China Center for Type Culture Collection (Wuhan, China) and named E. coli ETEC BE311 (CCTCC NO: M2019733).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Whole Genome Sequencing and CRISPR/Cas9 Gene Editing of Enterotoxigenic Escherichia coli BE311 for Fluorescence Labeling and Enterotoxin Analyses

doi: 10.3390/ijms23147502

Figure Lengend Snippet: Identification and characterization of E. coli BE311. ( A ), The expression of pQE30-GFP in different E. coil strains. ( B ), PCR amplification of the 16S rRNA of E. coli BE311. ( C ), Identification of the enterotoxins in E. coli BE311 by agarose gel electrophoresis (AGE). ( D ), Identification of the adhesions in E. coli BE311 by AGE.

Article Snippet: The BE311 strain was deposited in the China Center for Type Culture Collection (Wuhan, China) and named E. coli ETEC BE311 (CCTCC NO: M2019733).

Techniques: Expressing, Amplification, Agarose Gel Electrophoresis

Journal: International Journal of Molecular Sciences

Article Title: Whole Genome Sequencing and CRISPR/Cas9 Gene Editing of Enterotoxigenic Escherichia coli BE311 for Fluorescence Labeling and Enterotoxin Analyses

doi: 10.3390/ijms23147502

Figure Lengend Snippet: Whole genome sequencing of E. coli BE311. ( A ), The circus plot of E. coli BE311 genome. ( B ), The COG analysis of E. coli BE311 genome. ( C ), GO analysis of E. coli BE311 genome. ( D ), KEGG analysis of E. coli BE311 genome.

Article Snippet: The BE311 strain was deposited in the China Center for Type Culture Collection (Wuhan, China) and named E. coli ETEC BE311 (CCTCC NO: M2019733).

Techniques: Sequencing

Journal: International Journal of Molecular Sciences

Article Title: Whole Genome Sequencing and CRISPR/Cas9 Gene Editing of Enterotoxigenic Escherichia coli BE311 for Fluorescence Labeling and Enterotoxin Analyses

doi: 10.3390/ijms23147502

Figure Lengend Snippet: Expression and analysis of mCherry in BE311. ( A ), The process of knocking mCherry into BE311 based on CRISPR/Cas9. ( B ), Agarose gel electrophoresis detection of the mCherry in BE311 genome. ( C ), Red fluorescence expression in different strains with or without lactose inducement. ( D ), Stability analysis of BE311–mCherry–pQE30 with or without ampicillin. ( E ), Construction of the correlation curve between fluorescence intensity and concentration of BE311–mCherry–pQE30. ( F ), The fluorescence of E. coli BE311–mCherry–pQE30 captured under fluorescence microscope. ( G – I ), The gut microbiota of E. coli BE311–mCherry–pQE30-challenged mice was obtained and cultured in LB plate for 24 h. The fluorescence of colonies and bacteria in intestines ( H , I ) was identified under blue exciting light and fluorescence microscope. ( J – L ), The expression of IL-6, IL-8, and TNF-α in different intestines of E. coli BE311–mCherry–pQE30-challenged SD rats. Statistical significance was determined using Students’ t test. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The BE311 strain was deposited in the China Center for Type Culture Collection (Wuhan, China) and named E. coli ETEC BE311 (CCTCC NO: M2019733).

Techniques: Expressing, CRISPR, Agarose Gel Electrophoresis, Fluorescence, Concentration Assay, Microscopy, Cell Culture, Bacteria

Journal: International Journal of Molecular Sciences

Article Title: Whole Genome Sequencing and CRISPR/Cas9 Gene Editing of Enterotoxigenic Escherichia coli BE311 for Fluorescence Labeling and Enterotoxin Analyses

doi: 10.3390/ijms23147502

Figure Lengend Snippet: Construction and analysis of BE311-ST KO . ( A ), The knock-out process of ST gene in BE311. ( B ), Amplification of ST gene in wild-type and ST-knock-out BE311 strains. ( C ), The growth curve of BE311 and BE311-ST KO strains within 24 h. ( D ), IPEC-J2 cells were challenged with BE311 or BE311-ST KO for 6 h. Then the cellular apoptosis was examined by flow cytometry. ( E ), The total apoptosis rate in IPEC-J2 cells challenged with BE311 or BE311-ST KO for 6 h. ** p < 0.01, ns, no significant difference.

Article Snippet: The BE311 strain was deposited in the China Center for Type Culture Collection (Wuhan, China) and named E. coli ETEC BE311 (CCTCC NO: M2019733).

Techniques: Knock-Out, Amplification, Flow Cytometry

Journal: International Journal of Molecular Sciences

Article Title: Whole Genome Sequencing and CRISPR/Cas9 Gene Editing of Enterotoxigenic Escherichia coli BE311 for Fluorescence Labeling and Enterotoxin Analyses

doi: 10.3390/ijms23147502

Figure Lengend Snippet: Primers used in the construction of mCherry-labeled E. coli BE311.

Article Snippet: The BE311 strain was deposited in the China Center for Type Culture Collection (Wuhan, China) and named E. coli ETEC BE311 (CCTCC NO: M2019733).

Techniques: Sequencing