Techniques in Quantification and Localization of Gene Expression

1. Front Matter

   Pages i-xvii

   PDF

2. Basic Methods

   1. Front Matter

      Pages 1-1

      PDF

   2. Basic Flow Cytometry Instrumentation

      • Charles L. Goolsby, Cathy James

      Pages 3-22

   3. Fixation for In Situ Molecular Analysis

      • Bruce K. Patterson, Daniel K. Jiyamapa

      Pages 23-34

   4. A Method for Reduction of Green Wavelength Autofluorescence Emission in Cellular Preparations

      • Victoria L. Mosiman

      Pages 35-43

   5. Gene Quantification: Choosing the Target

      • Bruce K. Patterson

      Pages 45-53

   6. Detection and Quantification of Cytokine-Producing Cells by Immunostaining

      • Ulf Andersson, Mark J. Litton, Tom E. Fehniger, Ann-Kristin Ulfgren, Jan Andersson

      Pages 55-79

   7. In Situ Hybridization Using the bDNA Technology

      • Vincent P. Antao, Audrey N. Player, Janice A. Kolberg

      Pages 81-93

3. Specific Applications

   1. Front Matter

      Pages 95-95

      PDF

   2. Kaposi’s Sarcoma—Associated Human Herpes Virus Eight: Localization of Viral Gene Expression and Viral Latency

      • Scott J. Brodie, John N. Krieger, Catherine Diamond, Kurt Diem, Lawrence Corey

      Pages 97-106

   3. Pathogenetic Mechanisms of Animal Orbiviruses That Cause Disease at Low Copy Number

      • Scott J. Brodie, Patricia M. O’Hearn, Kurt Diem, David Muthui

      Pages 107-121

   4. Localization of Hepatitis C Virus Using Reverse Transcriptase Fluorescence In Situ 5′ Nuclease Assay

      • Bruce K. Patterson

      Pages 123-134

   5. Simultaneous Quantification and Localization of HIV gag-pol Expression Using Ultrasensitive Fluorescence In Situ Hybridization

      • Bruce K. Patterson

      Pages 135-142

   6. Quantification and Localization of Gene Expression: Future Directions

      • Bruce K. Patterson

      Pages 143-146

4. Back Matter

   Pages 147-150

   PDF

Can the son or daughter of a baseball pitcher or cricket bowler throw a ball 100 miles an hour? Is the son or daughter of an opera singer also an opera singer? Is a house with functional light switches lit? The line of thinking in these rhetorical questions also applies to human genetics. What do baseball pitchers, opera sing­ ers, light switches, and the Human Genome Project have in common? These questions address the issue of potential versus realization of function. Although sons and daughters of baseball pitchers and opera singers may have inherited the mechanical attributes to be baseball pitchers and opera singers, they may not, at any point in time, be baseball pitchers or opera singers. A house with functional light switches is not lit unless the light switches are on. Similarly, all of the genes discovered and sequenced as a result of the Human Genome Project are not expressed at the same time. Genome project information will allow us to deter­ mine the repertoire of genes in an individual, which is analogous to determining where the light switches in a house are located and whether they are functional (a mutation or deletion in the Genome Project Model). The pattern of "on" light switches in a house gives us functional information as to what the family inside is doing (e. g. , eating, reading, sleeping). Similarly, the pattern of gene expression (RNA) gives us information on what our bodies are doing (e. g.

• DNA
• HIV
• RNA
• biotechnology
• cell lines
• cytokine
• fluorescence
• gene expression
• genes
• genetics
• hybridization
• molecular biology
• protein
• tissue
• virus

• Laboratory of Viral Pathogenesis, Department of Pediatrics, Division of Special Infectious Diseases, Children’s Memorial Hospital, Chicago, USA

  Bruce K. Patterson

Policies and ethics