THE IN VITRO SYNTHESIS OF A NONINFECTIOUS COMPLEX CONTAINING BIOLOGICALLY ACTIVE VIRAL RNA

Abstract

The early events in the synthesis of infectious RNA by purified QB-replicase were analyzed. A latent period is accompanied by an eclipse of the initiating RNA templates as plaque-forming units. The latent period ends at about 6 min., the time required to complete the 1st new infectious strands. The eclipse is accompanied by a noninfectious complex with a sedimentation coefficient of 15S con-taining H3 isotope of the template and P32 which identified the product. The complex initiates infectious plus strands on heat denaturation. Massive formation of the complex requires freshly prepared viral RNA and replicase devoid of detectable RNase. This complex can be synthesized in any quantity with identifying isotopic labeling in its components making it possible to obtain the information required to define its structure and function.