Drescher B, Möhring R, Riedel H, Heider G
Arch Exp Veterinarmed. 1979;33(3):403-10.
RNA-directed DNA polymerase was isolated from the liver, spleen, and myeloblasts of chickens that had been infected with virus of avian myeloblastosis. The enzyme was chromatographically purified from the myeloblasts and brought to about 1,000-fold concentration. The method consisted in cell fractionation, lysis of the microsomal fraction, chromatography on Sephadex G-200 and phosphocellulose, as well as ultracentrifugation in the glycerol gradient. The cellular DNA polymerases alpha and beta were clearly separated from the DNA polymerase of avian myeloblastosis virus and could be distinguished from one another by template-specific reactions. The viral DNA polymerase activities in the microsomal fractions of liver, spleen, and myeloblasts were compared with one another. The amount of avian myeloblastosis virus and related DNA polymerase recorded from the myeloblasts was about six times that in the liver and four times higher than that in the spleen. The procedure described, together with the use of cell fractionation and gel filtration, is an appropriate method for biochemical detection of avian oncorna viruses in cells.
从感染了禽成髓细胞瘤病毒的鸡的肝脏、脾脏和成髓细胞中分离出了RNA指导的DNA聚合酶。该酶从成髓细胞中经色谱法纯化，并浓缩至约1000倍。方法包括细胞分级分离、微粒体部分的裂解、在葡聚糖凝胶G - 200和磷酸纤维素上的色谱分析以及在甘油梯度中的超速离心。细胞DNA聚合酶α和β与禽成髓细胞瘤病毒的DNA聚合酶明显分离，并且可以通过模板特异性反应相互区分。比较了肝脏、脾脏和成髓细胞微粒体部分中的病毒DNA聚合酶活性。从成髓细胞中记录到的禽成髓细胞瘤病毒和相关DNA聚合酶的量约为肝脏中的六倍，比脾脏中的高四倍。所述程序连同细胞分级分离和凝胶过滤的使用，是细胞中禽肿瘤病毒生化检测的合适方法。
