Yersinia Pestis Protocols

Dr. Ruifu Yang is a professor at the Beijing Institute of Microbiology and Epidemiology and the Chief-Scientist for National Basic Research Program (973 Program). He serves as the vice president for the Chinese Society for Microbiology. His research focuses on bacterial pathogenesis using genomics- and proteomics-based techniques. He has published more than 100 articles on international well-known journals, such as Nature Genetics, PNAS, BMC Microbiology.

• Isolation and identification of Yersinia pestis.- Isolation and characterization of Yersinia pestis bacteriophages.- Extraction of chromosomal DNA and plasmid from Yersinia pestis.- Extraction of total RNAs from Yersinia pestis.- Genotyping of Yersinia pestis.- Genome-wide variation analysis of Yersinia pestis.- Genome-wide detection of expressed sRNAs in Y. pestis with RNA-seq.- Validation of Y. pestis sRNAs with nonradiative-labeled Northern Blot.- Determination of transcriptional start sites (TSSs) in Y. pestis with a primer extension assay.- Purification of Hfq-associated RNAs with RNA immunoprecipitation (RIP).- Overexpression of target sRNA in tightly controlled multicopy plasmids.- Target validation of sRNA with a GFP reporter gene fusion system.- Inducible gene expression in Yersinia pestis.- Analysis of Yops secretion by Y. pestis Type III secretion system.- Co-Immunoprecipitation analysisfor detection of protein-protein interaction in Y. pestis.- Detection of type III secretion system needle assembly in Yersinia pestis by cross-linking of YscF.- Evaluation of protein translocation into eukaryotic cells by Y. pestis.- Adhesion analysis of Y. pestis to the host cells.- Construction of gene deletion mutants in Yersinia pestis.- CRISPR-Cas12a–assisted recombineering in Yersinia pestis.- Plasmid curing in Yersinia pestis.- Genetic regulation analysis of Yersinia pestis.- Investigation of Biofilm formation in Y. pestis.- Extraction and purification of F1 capsule antigen from Y. pestis.- Screening of regulator responses to Yersinia pestis infection with a concatenated tandem array of transcription factor response elements-pull down.- Animal models for Yersinia pestis.- Antibiotic resistance evaluation of Yersinia pestis.- PCR for detection and identification of Yersinia pestis.- Immunological methods for detection and identification of Yersinia pestis.- Analysis of fatty acids in Yersinia pestis by gas chromatography.- Evaluation of efficacy of Yersinia pestis  vaccine.- Online resources for Yersinia pestis.- Effects of Y. pestis on macrophage.