Sprague J, Condra J H, Arnheiter H, Lazzarini R A
J Virol. 1983 Feb;45(2):773-81. doi: 10.1128/JVI.45.2.773-781.1983.
A cDNA clone containing the entire vesicular stomatitis virus nucleocapsid gene was assembled by fusing portions of two partial clones. When the cDNA clone was inserted into a new general-purpose eucaryotic expression vector and introduced into appropriate host cells, abundant N-protein synthesis ensued. The expressed protein was indistinguishable from authentic N protein produced during vesicular stomatitis virus infections. The recombinant N protein was recognized by a polyclonal antibody and two different monoclonal antibodies and could not be resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis from authentic N. Our results suggest that the recombinant N protein produced in transfected cells rapidly aggregates into high-molecular-weight complexes in the absence of vesicular stomatitis virus genomic RNA.
通过融合两个部分克隆的片段，构建了一个包含水泡性口炎病毒核衣壳基因全长的cDNA克隆。当将该cDNA克隆插入到一个新的通用真核表达载体中并导入合适的宿主细胞后，大量的N蛋白得以合成。所表达的蛋白与水泡性口炎病毒感染期间产生的天然N蛋白无法区分。重组N蛋白可被多克隆抗体和两种不同的单克隆抗体识别，并且在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中无法与天然N蛋白区分开来。我们的结果表明，在没有水泡性口炎病毒基因组RNA的情况下，转染细胞中产生的重组N蛋白会迅速聚集成高分子量复合物。
