Spector D H, Varmus H E, Bishop J M
Proc Natl Acad Sci U S A. 1978 Sep;75(9):4102-6. doi: 10.1073/pnas.75.9.4102.
We have detected nucleotide sequences related to the transforming gene of avian sarcoma vius (ASV) in the DNA of uninfected vertebrates. Purified radioactive DNA (cDNAsarc) complementary to most of all of the gene (src) required for transformation of fibroblasts by ASV was annealed with DNA from a variety of normal species. Under conditions that facilitate pairing of partially matched nucleotide sequences (1.5 M NaCl, 59 degrees), cDNAsarc formed duplexes with chicken, human, calf, mouse, and salmon DNA but not with DNA from sea urchin, Drosophila, or Escherichia coli. The kinetics of duplex formation indicated that cDNAsarc was reacting with nucleotide sequences present in a single copy or at most a few copies per cell. In contrast to the preceding findings, nucleotide sequences complementary to the remainder of the ASV genome were observed only in chicken DNA. Thermal denaturation studies of the duplexes formed with cDNAsarc indicated a high degree of conservation of the nucleotide sequences related to src in vertebrate DNAs; the reductions in melting temperature suggested about 3--4% mismatching of cDNAsarc with chicken DNA and 8--10% mismatching of cDNAsarc with the other vertebrate DNAs.
我们在未感染的脊椎动物的DNA中检测到了与禽肉瘤病毒(ASV)转化基因相关的核苷酸序列。用与ASV转化成纤维细胞所需的大部分或全部基因(src)互补的纯化放射性DNA(cDNAsarc)与来自多种正常物种的DNA进行退火处理。在有利于部分匹配核苷酸序列配对的条件下(1.5M NaCl,59摄氏度),cDNAsarc与鸡、人、小牛、小鼠和鲑鱼的DNA形成双链体,但不与海胆、果蝇或大肠杆菌的DNA形成双链体。双链体形成的动力学表明,cDNAsarc正在与每个细胞中以单拷贝或最多几个拷贝存在的核苷酸序列发生反应。与之前的发现相反,仅在鸡的DNA中观察到与ASV基因组其余部分互补的核苷酸序列。对与cDNAsarc形成的双链体进行的热变性研究表明,脊椎动物DNA中与src相关的核苷酸序列高度保守;解链温度的降低表明cDNAsarc与鸡DNA约有3 - 4%的错配,与其他脊椎动物DNA约有8 - 10%的错配。