Isolation of a foot-and-mouth disease polyuridylic acid polymerase and its inhibition by antibody
Isolation of a foot-and-mouth disease polyuridylic acid polymerase and its inhibition by antibody
复制标题
口蹄疫多聚尿苷酸聚合酶的分离及其抗体抑制
作者：
J. Polatnick
从口蹄疫病毒感染的BHK细胞中分离到一种模板依赖的多聚尿苷酸[poly（U）]聚合酶。通过用30至50%饱和硫酸铵沉淀来浓缩细胞质提取物的20，000 x g上清液中的酶活性。通过十二烷基硫酸钠和1，1，2-三氯三氟乙烷提取使聚（U）聚合酶从膜上游离，并通过用2 M LiCl沉淀除去RNA。溶解的poly（U）聚合酶需要聚腺苷酸作为模板与寡尿苷酸引物和Mg 2+复合以获得活性，但被Mn 2+抑制。来自感染FMDV的动物的抗血清先前已显示抑制与内源RNA模板复合的FMDV RNA复制酶的活性。该抗血清还抑制poly（U）聚合酶的活性。通过与FMDV的病毒感染相关抗原吸收而耗尽抗体的抗血清不再抑制复制酶和聚合酶活性。证据表明，口蹄疫病毒RNA复制酶，聚（U）聚合酶，和病毒感染相关抗原共享一个共同的蛋白质。
A template-dependent polyuridylic acid [poly(U)] polymerase has been isolated from BHK cells infected with foot-and-mouth disease virus (FMDV). Enzyme activity in a 20,000 x g supernatant of a cytoplasmic extract was concentrated by precipitation with 30 to 50% saturated ammonium sulfate. The poly(U) polymerase was freed of membranes by sodium dodecyl sulfate and 1,1,2-trichlorotrifluoroethane extraction, and RNA was removed by precipitation with 2 M LiCl. The solubilized poly(U) polymerase required polyadenylic acid as template complexed to an oligouridylic acid primer and Mg2+ for activity, but was inhibited by Mn2+. Antisera from animals infected with FMDV had previously been shown to inhibit the activity of FMDV RNA replicase complexed to the endogenous RNA template. The same antisera also inhibited the activity of poly(U) polymerase. Antisera depleted of antibody by absorption with the virus infection-associated antigen of FMDV no longer inhibited replicase and polymerase activities. The evidence suggests that FMDV RNA replicase, poly(U) polymerase, and the virus infection-associated antigen share a common protein.