Tannock G A
Virology. 1977 May 15;78(2):500-10. doi: 10.1016/0042-6822(77)90126-x.
A homogeneous RNA complex with a sedimentation coefficient of 70 S and an apparent molecular weight of approximately 6.1 × 10 was released from purified P-labeled, mouse-brain-derived OC-43 virus after treatment with 1% sodium dodecyl sulfate (SDS) for 15 min at 23°. The complex was highly susceptible to heat, releasing 4 S RNA fragments at 37° and breaking down to fragments of 4–70 S at 60°; it was also degraded by centrifugation through dimethyl sulfoxide gradients. Unlike tobacco mosaic virus or Rous sarcoma virus RNA, OC-43 RNA prepared by extraction with phenol-SDS or phenol-chloroform degraded into a range of fragments with coefficients of 15–55 S; 4 S RNA was also present as a minor component. This suggests that (a) extensive nicking of a large RNA molecule has occurred during viral growth, due to ribonucleases which are inactivated during phenol extractions; (b) heterogeneity for OC-43 RNA is not due to internal ribonuclease activity and fragments are held together by noncovalent linkages much weaker than those present in the 70 S retroviral RNA complex, or by small proteins; or, most probably, (c) a combination of extensive nicking and weak noncovalent linkages results in the heterogeneous denaturation products.
用1%十二烷基硫酸钠(SDS)在23°处理15分钟后,从纯化的经P标记、源自小鼠脑的OC - 43病毒中释放出一种沉降系数为70 S、表观分子量约为6.1×10的均匀RNA复合物。该复合物对热高度敏感,在37°时释放出4 S RNA片段,在60°时分解为4 - 70 S的片段;通过二甲基亚砜梯度离心也会使其降解。与烟草花叶病毒或劳氏肉瘤病毒RNA不同,用苯酚 - SDS或苯酚 - 氯仿提取制备的OC - 43 RNA降解为一系列沉降系数为15 - 55 S的片段;4 S RNA也作为次要成分存在。这表明:(a)在病毒生长过程中,由于在苯酚提取过程中失活的核糖核酸酶,大RNA分子发生了广泛的切口;(b)OC - 43 RNA的异质性不是由于内部核糖核酸酶活性,片段通过比70 S逆转录病毒RNA复合物中存在的非共价键弱得多的非共价键或小蛋白质结合在一起;或者,最有可能的是,(c)广泛的切口和弱非共价键的组合导致了异质的变性产物。