Assessment of fHbp Expression Level by Reverse Transcriptase Quantitative PCR and Promoter Sequence Analysis
Reverse transcriptase quantitative PCR (RT qPCR) is widely used for assessing the levels of expression of specific genes in various organisms. Here we describe a RT qPCR assay designed to determine the level of expression of fHbp in multiple isolates of Neisseria meningitidis. The level of expression is measured by a two-step qPCR and is associated with a promoter region analysis.
Key wordsGene expression fHbp qPCR Intergenic region Reverse transcription
- 5.Cayrou C, Akinduko AA, Mirkes EM, Lucidarme J, Clark SA, Green LR et al (2018) Clustered intergenic region sequences as predictors of factor H binding protein expression patterns and for assessing Neisseria meningitidis strain coverage by meningococcal vaccines. PLoS One 13(5):e0197186CrossRefGoogle Scholar
- 6.Higuchi R, Fockler C, Dollinger G, Watson R (1993) Kinetic PCR analysis: real-time monitoring of DNA amplification reactions. Biotechnology (N Y) 11:1026–1030Google Scholar
- 7.Sanders H, Brehony C, Maiden MC, Vipond C, Feavers IM (2012). The effect of iron availability on transcription of the Neisseria meningitidis fHbp gene varies among clonal complexes. Microbiology 158:869–876Google Scholar