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November 18, 1974

Ribonucleic Acid: I. The Test-Tube Synthesis of a Viral Nucleic Acid II. The Development and Use of Molecular Hybridization

Author Affiliations

From the Institute of Cancer Research, and Department of Human Genetics and Development, College of Physicians and Surgeons, Columbia University, New York.

JAMA. 1974;230(7):1036-1042. doi:10.1001/jama.1974.03240070068041

Two sets of investigations are briefly described. The first is aimed at an understanding of the molecular life history of lytic RNA virus with particular emphasis on the mechanism of RNA replication. The studies resulted in the isolation of the relevant RNA-directed RNA polymerase (replicase) and in the demonstration that this enzyme would mediate the continuous test-tube synthesis of unlimited quantities of infectious and biologically competent viral nucleic acid. This unique situation was exploited to examine the extracellular mutation and evolution of replicating molecules. The investigation culminated recently with the complete sequence determination of one of the mutant RNA molecules, an achievement that identified the secondary and tertiary structures as the source of the phenotypic wealth required to explain the evolutionary potential of these molecules.

The second investigation began with an attempt to provide a rigorous proof that DNA communicated its instructions via RNA complementary transcript. A procedure for annealing RNA to its complementary DNA and a sensitive method for detecting the resulting hybrid complexes were developed. Molecular hybridization thus provided the first direct evidence supporting the concept of cellular RNA as genetic messages. We further refined and simplified the procedure in an attempt to convert it into a powerful and flexible tool. RNA-DNA hybridization was then successfully used in the resolution of many of the central problems of molecular genetics. In this manner, nucleic acid hybridization became one of the most widely used molecular biologic techniques of the last decade. Its recent introduction into the cancer problem has provided the first molecular evidence of tumor viral information in human neoplasias.

(JAMA 230:1036-1042, 1974)