Molecular cloning of two paralytogenic, temperature-sensitive mutants, ts1 and ts7, and the parental wild-type Moloney murine leukemia virus

Abstract

ts1 and ts7, the paralytogenic, temperature-sensitive mutants of Moloney murine leukemia virus (MoMuLV), together with their wild-type parent, MoMuLV-TB, were molecularly cloned. ts1-19, ts7-22, and wt-25, the infectious viruses obtained on transfection to NIH/3T3 cells of the lambda Charon 21A recombinants of ts1, ts7, and wt, were found to have retained the characteristics of their non-molecularly cloned parents. In contrast to the wt virus, ts1-19 and ts7-22 are temperature-sensitive, inefficient in the intracellular processing of Pr80env at the restrictive temperature, and able to induce paralysis in CFW/D mice. Like the non-molecularly cloned ts7, the ts7-22 virion was also shown to be heat labile. The heat lability of the ts7 virion distinguishes it from ts1. Endonuclease restriction mapping with 11 endonucleases demonstrated that the base composition of MoMuLV-TB differs from that of the standard MoMuLV, but no difference was detected between the molecularly cloned ts1 and ts7 genomes. However, ts1 and ts7 differ from MoMuLV in the loss or acquisition of four different restriction sites, whereas they differ from MoMuLV-TB in the loss or acquisition of three different restriction sites.