Chirikjian J G, Rye L, Papas T S
Proc Natl Acad Sci U S A. 1975 Mar;72(3):1142-6. doi: 10.1073/pnas.72.3.1142.
Pyran covalently linked to cyanogen bromide-activated Sepharose has been shown to be an effective affinity matrix for several viral DNA polymerases. Differential salt elution of viral compared with cellular polymerases, as well as substrate elution, suggests the affinity nature for the matrix. Unlike some other affinity systems described, pyran-Sepharose is totally resistant to nuclease digestion and is stable at 4 degrees for several months. DNA polymerases isolated from several viruses by detergent treatment were recovered in good yield. Analysis of iodinated proteins by sodium dodecyl sulfate-gel electrophoresis revealed that the DNA polymerase of avian myeloblastosis virus found in crude preparations of the virus could be purified nearly to homogeneity by a single passage through the column. These results suggest that pyran-Sepharose is an effective affinity column that is potentially adaptable as part of a general purification procedure for viral DNA polymerases.
已证明与溴化氰活化的琼脂糖共价连接的吡喃是几种病毒DNA聚合酶的有效亲和基质。与细胞聚合酶相比，病毒聚合酶的差异盐洗脱以及底物洗脱表明了该基质的亲和性质。与所描述的其他一些亲和系统不同，吡喃 - 琼脂糖对核酸酶消化完全抗性，并且在4℃下稳定数月。通过去污剂处理从几种病毒中分离的DNA聚合酶以良好的产率回收。通过十二烷基硫酸钠 - 凝胶电泳对碘化蛋白质的分析表明，在病毒粗制品中发现的禽成髓细胞瘤病毒的DNA聚合酶通过单次通过该柱可纯化至几乎同质。这些结果表明，吡喃 - 琼脂糖是一种有效的亲和柱，有可能作为病毒DNA聚合酶通用纯化程序的一部分。
