Shtivelman E, Zakut R, Canaani E
Proc Natl Acad Sci U S A. 1984 Jan;81(2):294-8. doi: 10.1073/pnas.81.2.294.
Nonproducer transformants infected with wild-type Moloney murine sarcoma virus were screened for the generation of variants with reorganized genomes. Seven of 20 lines contained such viral genomes, 5 of which were found to be nonrescuable. Blotting analysis indicated that viral RNA molecules transcribed from these variant genomes could not be encapsidated into virions. The nonrescuable genomes were molecularly cloned and all were found to have suffered deletions or deletions/inversions involving the 5' long terminal repeat as well as some adjacent sequences. Nucleotide sequence analysis suggested that the long terminal repeat or the tetranucleotides G-G-T-C and G-A-C-C (or both) were involved in the generation of these mutants. Transfection studies showed that the cloned DNAs of the 5 mutants transformed NIH/3T3 monolayers. Removal of the 3' long terminal repeat from the genomes that lacked the 5' long terminal repeat or carried it in an inverted orientation abolished or considerably reduced the transforming activity.
对感染野生型莫洛尼氏鼠肉瘤病毒的无病毒产生转化体进行筛选，以寻找基因组发生重组的变体。20个品系中有7个含有此类病毒基因组，其中5个被发现无法拯救。印迹分析表明，从这些变体基因组转录的病毒RNA分子无法包装到病毒粒子中。对无法拯救的基因组进行分子克隆，发现它们都发生了缺失或涉及5'长末端重复序列以及一些相邻序列的缺失/倒位。核苷酸序列分析表明，长末端重复序列或四核苷酸G-G-T-C和G-A-C-C（或两者）与这些突变体的产生有关。转染研究表明，5个突变体的克隆DNA可转化NIH/3T3单层细胞。从缺乏5'长末端重复序列或以反向携带该序列的基因组中去除3'长末端重复序列，可消除或显著降低转化活性。
