Preparation of RNA from bacteria infected with bacteriophages: a case study from the marine unicellular Synechococcus sp. WH7803 infected by phage S-PM2.
Bacteriophages manipulate bacterial gene expression in order to express their own genes or influence bacterial metabolism. Gene expression can be studied using real-time PCR or microarrays. Either technique requires the prior isolation of high quality RNA uncontaminated by the presence of genomic DNA. We outline the considerations necessary when working with bacteriophage infected bacterial cells. We also give an example of a protocol for extraction and quantification of high quality RNA from infected bacterial cells, using the marine cyanobacterium WH7803 and the phage S-PM2 as a case study. This protocol can be modified to extract RNA from the host/bacteriophage of interest.
Shan
Jinyu
J
Department of Biological Sciences, University of Warwick, Coventry, UK.
Clokie
Martha
M
D001435:Bacteriophages / Q000235:genetics* / Q000502:physiology; D054884:Host-Pathogen Interactions; D012329:RNA, Bacterial / Q000235:genetics* / Q000302:isolation & purification; D012367:RNA, Viral / Q000235:genetics* / Q000302:isolation & purification; D046940:Synechococcus / Q000235:genetics* / Q000821:virology
D012329:RNA, Bacterial; D012367:RNA, Viral