Transformation of competent cells

Growth and preparation of Streptococcus pneumoniae

Making sheep’s blood agar plates for culture of Streptococcus pneumoniae

Intranasal colonization of Streptococcus pneumoniae (and measurement of gene expression and leukocyte recruitment to the nasopharnyx)

• This is a detailed protocol for preparing Streptococcus pneumoniae and intranasal colonization of mice. Puchta A, Verschoor CP, Thurn T, Bowdish DM. Characterization of inflammatory responses during intranasal colonization with Streptococcus pneumoniae. J Vis Exp. 2014 Jan 17;(83):e50490. doi: 10.3791/50490. For access to an accompanying video, please contact Dr. Bowdish (bowdish@mcmaster.ca)
• We have a protocol specific to TIGR4/P1542 here.
• Also used in: Thevaranjan N, et al. Streptococcus pneumoniae Colonization Disrupts the Microbial Community within the Upper Respiratory Tract of Aging Mice.Infect Immun. 2016 Mar 24;84(4):906-16. doi: 10.1128/IAI.01275-15. 

Pneumolysin purification from an expression plasmid.

CFSE labelling of S pneumoniae

• This protocol uses CFSE to label S. pneumoniae for flow cytometry or microscopy experiments.

Pneumococcal Cell Wall Purification

• This protocol describes how to purify the pneumococcal cell wall. The purified preparation will not contain proteins but will contain the peptidoglycan layer and teichoic acids.

Phenol-sulfuric acid Mannose detection assay.

• This assay is sensitive to mannose from 1 – 100 𝜇g but will also detect other carbohydrates. We use this assay to quantify of complex carbohydrates in the bacterial cell wall