Oskarsson M K, Elder J H, Gautsch J W, Lerner R A, Vande Woude G F
Proc Natl Acad Sci U S A. 1978 Oct;75(10):4694-8. doi: 10.1073/pnas.75.10.4694.
The gene order of the ml Moloney sarcoma virus (mlMSV) specific pP60gag (P60) was determined by direct chemical analysis of the polyprotein. P60 was cleaved with cyanogen bromide (CNBr) into eight partial and complete fragments ranging in mass from 10,000 daltons to 58,000 daltons. Peptide maps of these fragments were compared to maps of p15, p12, and three CNBr fragments of p30. The polarity of p15 and p12 in a CNBr fragment of P60 was determined by carboxypeptidase A digestion; likewise the CNBr fragments of p30 were ordered by aminopeptidase digestion. The linear arrangement of P60 CNBr fragments gave the gene order of NH2-p15-p12-p30-COOH. The m3 isolate of MSV expresses a P70 gag polyprotein. Peptide maps of 48,000-dalton CNBr fragments of m3 P70 and ml P60 were similar and suggested that both polyproteins were similar through the NH2-terminal two-thirds of p30. However, the presence of peptides unique to the 10,500-dalton COOH-terminal fragment of m1MSV p30 and not present in the p30 of either m3MSV or Moloney leukemia virus suggested that the gag gene deletion in the m1 isolate begins in the p30 reading frame.
通过对多聚蛋白进行直接化学分析，确定了莫洛尼氏肉瘤病毒（mlMSV）特异性pP60gag（P60）的基因顺序。用溴化氰（CNBr）将P60切割成八个部分片段和完整片段，质量范围从10,000道尔顿到58,000道尔顿。将这些片段的肽图与p15、p12以及p30的三个CNBr片段的图谱进行比较。通过羧肽酶A消化确定P60的CNBr片段中p15和p12的极性；同样，通过氨肽酶消化确定p30的CNBr片段的顺序。P60的CNBr片段的线性排列给出了NH2-p15-p12-p30-COOH的基因顺序。MSV的m3分离株表达一种P70 gag多聚蛋白。m3 P70和ml P60的48,000道尔顿CNBr片段的肽图相似，表明这两种多聚蛋白在p30的NH2末端三分之二区域相似。然而，m1MSV p30的10,500道尔顿COOH末端片段特有的肽段在m3MSV或莫洛尼氏白血病病毒的p30中不存在，这表明m1分离株中的gag基因缺失始于p30阅读框。
