A RAPID RT-PCR BASED METHOD TO ISOLATE COMPLEMENTARY-DNA FRAGMENTS FLANKING RETROVIRUS INTEGRATION SITES

Proto-oncogenes in retrovirally induced myeloid mouse leukemias are-fr equently activated following retroviral insertion. The identification; of common virus integration sites (VISs) and isolation of the transfo rming oncogene is laborious and time consuming. We established a rapid and simple PCR based procedure which facilitates the identification o f VISs and novel proto-oncogenes. Complementary DNA fragments adjacent to retrovirus integration sites were selectively isolated by applying a reverse transcriptase (RT) reaction using an oligo(dT)-adaptor prim er, followed by PCR using the adaptor sequence and a retrovirus long t erminal repeat (LTR) specific primer. Multiple chimeric cDNA fragments suitable for Southern and northern blot analysis were isolated.