Cytomegalovirus Protocols by John Sinclair

By John Sinclair

John Sinclair and a panel of specialist investigators current a finished number of mobile and molecular suggestions for the research of cytomegalovirus (CMV) biology and its pathogenetic mechanisms. The methods-all defined in step by step aspect with prepared reproducibility in mind-range from easy virus tradition to advanced molecular research of CMV constitution and serve as. incorporated are tools for CMV detection utilizing either immunological and organic innovations, equipment for studying primary points of the CMV an infection cycle, and techniques for reading T mobilephone reaction to cytomegalovirus an infection within the human host. finished and state of the art, Cytomegalovirus Protocols offers investigators with a set of the most important equipment which are illuminating not just the fundamental biology of this complicated and fascinating human herpesvirus, but in addition its major function in human infectious ailments and their emergent remedies.

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21. Boyle, K. A. and Compton, T. (1997) Receptor binding properties of a soluble form of the glycoprotein B of human cytomegalovirus. 22. Kari, B. and Gehrz, R. (1992) A human cytomegalovirus glycoprotein complex designated gC-II is a major heparin-binding component of the envelope. J. Virol. 66, 1761–1764. 23. Nowlin, D. , Cooper, N. , and Compton, T. (1991) Expression of a human cytomegalovirus receptor correlates with infectibility of cells. J. Virol. 65, 3114–3121. 24. Wright, J. , and Wasi, S.

HCMV Envelope Proteins Analysis of the nucleotide sequence of HCMV showed a total of 54 open reading frames (ORFs) that have characteristics of glycoprotein genes or of exons of glycoprotein genes (1). Some have homology to immunoglobulin superfamily proteins, others are multiple membrane spanning, and a few have recognized homology to glycoproteins of other herpesviruses. Three major glycoprotein complexes present in the envelope of HCMV have been biochemically characterized. It is probable that other glycoproteins are present in the virion envelope, but they remain uncharacterized and undetected.

Precipitate overnight at –20°C with equal volume isopropanol after extracting the DNase-free RNA with an equal volume of acidified phenol–chloroform–isoamyl alcohol supplied with the RNagents kit. 9. Resuspend the RNA pellet in an appropriate volume of RNase-free water, depending on the anticipated RNA concentration. Store at –70°C. 7. Reverse Transcription Controls for the reverse transcription reaction are provided by setting up an equal number of tubes substituting RNase-free water for Oligo (dT)12–18 primer, and in separate reactions, substituting RNase-free water for Superscript II reverse transcriptase.

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