Transcriptase activity associated with rabies virion
Transcriptase activity associated with rabies virion
复制标题
与狂犬病毒粒子相关的转录酶活性
作者：
A. Kawai
在体外研究了狂犬病病毒粒子相关的转录酶活性，并与水泡性口炎病毒的新泽西血清型进行了比较。影响[3 H]GMP进入酸不溶性物质的去污剂浓度对于两种病毒而言显著不同。水泡性口炎病毒新泽西需要0.05 - 0.1%的非离子去污剂，而狂犬病病毒粒子不能完全激活，除非使用4 - 5%的去污剂。其他最佳条件如下：40 mM NaCl、5 mM Mg 2+、40 mM Tris-盐酸盐（pH 7.4）、5 mM二硫苏糖醇和30 ℃。该反应需要四个核苷三磷酸。狂犬病病毒体酶的RNA合成的初始速率为每小时140 pmol GMP掺入/mg病毒蛋白，并线性增加，直到约8小时，具有轻微的初始滞后期。当狂犬病毒粒子在33 ℃下生长时，与L蛋白含量相关的酶活性最高。产物为单链RNA，其碱基序列与狂犬病毒RNA互补。大部分合成的RNA沉积在6- 16 S。
Rabies virion-associated transcriptase activity was investigated in vitro and compared with that of the New Jersey serotype of vesicular stomatitis virus. The concentration of detergent that affected [3H]GMP incoporation into acid-insoluble material was significantly different for both viruses. Vesicular stomatitis virus New Jersey required 0.05 to 0.1% nonionic detergent, whereas rabies virion could not be fully activated unless 4 to 5% detergent was used. Other optimal conditions were as follows: 40 mM NaCl, 5 mM Mg2+, 40 mM Tris-hydrochloride (pH 7.4), 5 mM dithiothreitol, and 30 degrees C. The reaction required four nucleoside triphosphates. The initial rate of RNA synthesis by rabies virion enzyme was 140 pmol of GMP incorporated/mg of viral protein per h and linearly increased until about 8 h, with a slight initial lag phase. The enzyme activity that correlated with the content of L protein was highest when rabies virions were grown at 33 degrees C. The product was single-stranded RNA, which was complementary in base sequences to rabies viral RNA. Most of the RNA synthesized sedimented at 6-16S.