Imblum R L, Wagner R R
J Virol. 1975 Jun;15(6):1357-66. doi: 10.1128/JVI.15.6.1357-1366.1975.
In search of an anti-transcriptase, antibody was raised in rabbits to partially purified, soluble NS protein present in cytoplasmic extracts of cells infected with the Indiana serotype of vesicular stomatitis (VSInd) virus. This antiserum gave specific reactions of identity by agar immunodiffusion with both cytoplasmic and virion NS protein. NS antiserum also preferentially precipitated NS 3-H-labeled protein from infected cytoplasmic extracts, whereas anti-whole VSInd virion serum also precipitated N 3-H-labeled protein from extracts both of infected cytoplasm and virion nucleocapsids. Transcriptase activity of VSInd cytoplasmic or virion-derived nucleocapsids was effectively inhibited by ribonuclease-free immunoglubulin prepared from homologous NSInd antiserum or from anti-whole vesicular stomatitis virus serum. Transcriptase activity of heterologous New Jersey serotype (VSNJ) nucleocapsids and virions was not appreciably affected by anti-NSInd or by anti-whole VSInd virion gamma globulin. Anti-NS gamma glubulin immediately switched off RNA synthesis by actively transcribing VSInd nucleocapsids, a finding which suggests that NS antibody inhibits RNA chain elongation.
为了寻找一种抗转录酶,用感染水泡性口炎病毒印第安纳血清型(VSInd)的细胞胞质提取物中存在的部分纯化的可溶性NS蛋白对家兔进行免疫,制备抗体。该抗血清通过琼脂免疫扩散与胞质和病毒粒子NS蛋白产生特异性的同一性反应。NS抗血清还优先从感染的胞质提取物中沉淀出NS 3-H标记的蛋白,而抗整个VSInd病毒粒子血清也从感染的胞质提取物和病毒粒子核衣壳提取物中沉淀出N 3-H标记的蛋白。从同源NSInd抗血清或抗整个水泡性口炎病毒血清制备的无核糖核酸酶免疫球蛋白有效地抑制了VSInd胞质或病毒粒子来源的核衣壳的转录酶活性。异源新泽西血清型(VSNJ)核衣壳和病毒粒子的转录酶活性不受抗NSInd或抗整个VSInd病毒粒子γ球蛋白的明显影响。抗NSγ球蛋白通过积极转录VSInd核衣壳立即关闭RNA合成,这一发现表明NS抗体抑制RNA链延伸。