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Human Genome Quarterly, Summer 1989; 1(2)
Electrophoretic Techniques in Molecular Biology
The 1989 Annual Meeting of the American Electrophoresis Society (AES) was held July 6-11 in Washington, D.C. The society, whose president is Carl Merrill of the National Institutes of Health, conducted its first annual meeting to facilitate communication among researchers who are using electrophoretic methods, theories, and applications. The program included a comparable number of presentations relating to both protein and DNA electrophoretic separations strategies and methodologies.
Some presentations relating to genome mapping and sequencing efforts are highlighted below:
Leroy Hood (California Institute of Technology) gave a comprehensive assessment of the technology developments needed to confront the challenges associated with genome mapping and sequencing. In a presentation that encompassed both protein and DNA separations/sequencing schemes, Hood offered that deciphering the encyclopedia of life-the genetic code-will increase understanding of the multiplicity of interrelated biochemical events that comprise normal life processes. Also discussed were direct applications of genome sequencing work to medical research of abnormal life processes, particularly in the area of preventive medicine.
Norman Anderson (Large Scale Biology Corp.) discussed the use of the protein gene product as a starting point to obtain the corresponding DNA and, ultimately, the DNA sequence; the potential problems of creating large-scale systems from smaller functional systems; the need for standardization of reagents; and the forces that influence funding agencies.
Charles Cantor (Lawrence Berkeley Laboratory) described some of the latest pulsed-field gel (PFG) electrophoresis developments in his laboratory. In a session devoted to PFG techniques, he said that because no one has made a detailed, quantitative description of the pulsed-field phenomenon, there are more opportunities for serendipitous discoveries. Since the publication of the PFG method in 1984 by Schwartz and Cantor (Cell 37:67), many variations and improvements have been made both in their laboratories and by others. With the advent of large-size standards, the implementation of the Smith-Birnstiel restriction mapping approaches, and the use of telomeric probes, large regions of human chromosomes can now be mapped efficiently by using PFG techniques.
David Patterson (Eleanor Roosevelt Institute for Cancer Research) described his work on mapping chromosome 21. He uses a modification of the PFG technique to gain a better understanding of the role that chromosome 21 trisomy plays in Down's Syndrome.
To illustrate his method of constructing computer models that demonstrate how DNA molecules travel through agarose gels, Steven Smith (University of Washington, Seattle) showed a dramatic videotape of stained DNA fragments (50 Kbp-1 Mbp), which were undergoing agarose gel electrophoresis. The process was filmed through a fluorescence microscope aided by an image intensifier. Using these video tapes, Smith is better able to model and understand the movement (and hence separation) of DNA molecules in PFG electrophoresis systems. A listing of his Pascal program source code for modeling movement of DNA through gels and a copy of the video tape can be obtained from the University of Washington.
Michael Harrington (California Institute of Technology) described improvements for implementation of the two-dimensional gel electrophoresis (2-DGE) technique both in the clinical laboratory and in the study of developmental biology systems. Starting with the 2-DGE analysis of gene products (proteins) allows identification of the genes that code for those proteins. Discussed by both Hood and Anderson earlier in the meeting, these genetics techniques are complementary to the direct mapping and DNA sequencing approach.
Terry Landers (Life Technologies) reviewed a variety of techniques for modifying nucleic acids by incorporating radioisotopes and non-isotopic reporter groups for detection.
Bruce Roe (University of Oklahoma) and Richard Wilson (California Institute of Technology) gave useful presentations concerning automated approaches to DNA sequencing.
Videotape Information:
Pascal Source Code Information: Internet: steve@snert.cpac.washington.edu
Submitted by: Betty K. Mansfield, HGMIS
The electronic form of the newsletter may be cited in the following style:
Human Genome Program, U.S. Department of Energy, Human Genome News (v1n2).
The Human Genome Project (HGP) was an international 13-year effort, 1990 to 2003. Primary goals were to discover the complete set of human genes and make them accessible for further biological study, and determine the complete sequence of DNA bases in the human genome. See Timeline for more HGP history.
Published from 1989 until 2002, this newsletter facilitated HGP communication, helped prevent duplication of research effort, and informed persons interested in genome research.
