Abstract

The general goal of this program is the development of new techniques to study macromolecules and assemblies. The current specific objectives are to develop methods for handling and analysis of chromosome-sized DNA molecules and methods for producing labeled proteins in vivo. Pulsed field gradient gel electrophoresis allows high resolution separations of DNA molecules as large as 5000 kb. This will be used to develop rapid gene isolataion and mapping methods. Refinements of current pulsed field techniques and methods for specific fragmentation of larger DNA will be developed so that the rapid mapping methods can be extended to human genes. Among the proposed applications are electrophoretic karyotypes and genome finger prints, techniques for exploring genome structure adjacent to a cloned marker, and methods for exploring the dynamics of mobile genetic elements. In other studies the genes for strepavidin and aequorin will be cloned and expressed in mammalian cells. Vectors will be constructed that produce fusions between these proteins and normal cellular proteins. This will allow specific in vivo labeling of any gene product.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM014825-21
Application #
3268693
Study Section
Molecular Biology Study Section (MBY)
Project Start
1975-02-01
Project End
1990-01-31
Budget Start
1987-02-01
Budget End
1988-01-31
Support Year
21
Fiscal Year
1987
Total Cost
Indirect Cost

  Institution

Name
Columbia University (N.Y.)
Department
Type
Schools of Medicine
DUNS #
064931884
City
New York
State
NY
Country
United States
Zip Code
10027

  Publications

Condemine, G; Smith, C L (1990) Transcription regulates oxolinic acid-induced DNA gyrase cleavage at specific sites on the E. coli chromosome. Nucleic Acids Res 18:7389-96
Banerjee, S; Cantor, C R (1990) Nucleosome assembly of simian virus 40 DNA in a mammalian cell extract. Mol Cell Biol 10:2863-73
Smith, C L; Cantor, C R (1989) Evolving strategies for making physical maps of mammalian chromosomes. Genome 31:1055-8
Fan, J B; Chikashige, Y; Smith, C L et al. (1989) Construction of a Not I restriction map of the fission yeast Schizosaccharomyces pombe genome. Nucleic Acids Res 17:2801-18
Cheng, J F; Smith, C L; Cantor, C R (1989) Isolation and characterization of a human telomere. Nucleic Acids Res 17:6109-27
Ohki, M; Smith, C L (1989) Tracking bacterial DNA replication forks in vivo by pulsed field gel electrophoresis. Nucleic Acids Res 17:3479-90
Mathew, M K; Smith, C L; Cantor, C R (1988) High-resolution separation and accurate size determination in pulsed-field gel electrophoresis of DNA. 2. Effect of pulse time and electric field strength and implications for models of the separation process. Biochemistry 27:9210-6
Mathew, M K; Hui, C F; Smith, C L et al. (1988) High-resolution separation and accurate size determination in pulsed-field gel electrophoresis of DNA. 4. Influence of DNA topology. Biochemistry 27:9222-6
Mathew, M K; Smith, C L; Cantor, C R (1988) High-resolution separation and accurate size determination in pulsed-field gel electrophoresis of DNA. 1. DNA size standards and the effect of agarose and temperature. Biochemistry 27:9204-10
Cantor, C R; Smith, C L; Mathew, M K (1988) Pulsed-field gel electrophoresis of very large DNA molecules. Annu Rev Biophys Biophys Chem 17:287-304

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