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Single Cell Papers
Sun Z, Chen L, Xin H, Jiang Y, Huang Q, Cillo AR, Tabib T, Kolls JK, Bruno TC, Lafyatis R, Vignali DAA, Chen K, Ding Y, Hu M, Chen W. A Bayesian mixture model for clustering droplet-based single-cell transcriptomic data from population studies. Nat Commun. 2019 Apr 9;10(1):1649.
Castillo-Dela Cruz P, Wanek AG, Kumar P, An X, Elsegeiny W, Horne W, Fitch A, Burr AHP, Gopalakrishna KP, Chen K, Methé BA, Canna SW, Hand TW, Kolls JK. Intestinal IL-17R Signaling Constrains IL-18-Driven Liver Inflammation by the Regulation of Microbiome-Derived Products. Cell Rep. 2019 Nov 19;29(8):2270-2283.
Iwanaga N, Sandquist I, Wanek A, McCombs J, Song K, Kolls JK. Host immunology and rational immunotherapy for carbapenem-resistant Klebsiella pneumoniae infection. JCI Insight. 2020 Apr 23;5(8):e135591.
Methanol Fixation of Cells for Single Cell RNA Sequencing
E. coli Expression of OmpX
OmpX sequence was obtained from NCBI website and DNA was PCR cloned using K. pneumoniae ATCC-43816 (KP-43816) cDNA as a template and sub-cloned into pET-DEST42 prokaryotic expression vector. BL21 competent E. coli was then transformed by pET-DEST42 plasmid DNA encoding OmpX under Ampicillin selection. PCR identification positive transformants were then grown in bulk culture and OmpX expression was induced by adding IPTG. OmpX expressing E. coli was then lysed in Solulyse buffer (Genlantis Inc.) and OmpX protein was identified by western blotting using anti-V5 antibody and purified by 6xHis tag chromatography using GraviTrap columns (GE healthcare).