Stewart W E, Sarkar F H, Taira H, Hall A, Nagata S, Weissmann C
Gene. 1980 Nov;11(3-4):181-6. doi: 10.1016/0378-1119(80)90058-x.
Human interferon (IFN) prepared from virus-induced human leukocyte suspensions (leukocyte-derived interferon) was compared to the IFN extracted from Escherichia coli harboring a human interferon-alpha cDNA hybrid plasmid (Hif-SN35-AH-L6). E coli-derived IFN was 20 to 50 times more active than leukocyte-derived IFN on heterologous bovine, feline, murine and guinea pig cells, relative to the activity on human cells. After partial purification by affinity chromatography on an anti-human lymphoblastoid IFN antibody column, the IFN was analyzed by SDS-polyacrylamide gel electrophoresis. While leukocyte-derived IFN gave a heterogeneous pattern with major peaks of activity of 24000 and 19000 daltons, E. coli-derived IFN gave a heterogeneous peak of activity at about 17-18000 daltons. The leading edge of leukocyte-derived IFN in SDS-polyacrylamide gels was significantly more active on bovine cells than on human cells and coincided in mobility with E. coli-derived IFN, which was also much more active on bone than on human cells. After reduction with mercaptoethanol in SDS, the E. coli-derived IFN lost no activity, whereas the leukocyte-derived IFN lost about 90% of its activity. After reduction, E. coli-derived IFN migrated in SDS-polyacrylamide gels as a single peak at 24000 daltons, as did the residual activity of reduced leukocyte-derived interferon. Out data suggest that the interferon produced by the E. coli harboring the clone Hif-SN35-AH-L6 is analogous in size and cross-species activity to one of the molecular species of leukocyte-derived interferon.
将从病毒诱导的人白细胞悬液中制备的人干扰素（IFN）（白细胞源性干扰素）与从携带人干扰素-α cDNA 杂交质粒（Hif-SN35-AH-L6）的大肠杆菌中提取的 IFN 进行了比较。相对于对人细胞的活性，大肠杆菌源性 IFN 对异源牛、猫、鼠和豚鼠细胞的活性比白细胞源性 IFN 高 20 至 50 倍。在抗人淋巴母细胞 IFN 抗体柱上进行亲和层析部分纯化后，通过 SDS-聚丙烯酰胺凝胶电泳对 IFN 进行分析。白细胞源性 IFN 呈现出异质性图谱，主要活性峰为 24000 和 19000 道尔顿，而大肠杆菌源性 IFN 在约 17000 - 18000 道尔顿处呈现出一个异质性活性峰。在 SDS-聚丙烯酰胺凝胶中，白细胞源性 IFN 的前沿在牛细胞上的活性明显高于在人细胞上的活性，并且其迁移率与大肠杆菌源性 IFN 一致，大肠杆菌源性 IFN 在牛细胞上的活性也远高于在人细胞上的活性。在 SDS 中用巯基乙醇还原后，大肠杆菌源性 IFN 没有失去活性，而白细胞源性 IFN 失去了约 90%的活性。还原后，大肠杆菌源性 IFN 在 SDS-聚丙烯酰胺凝胶中迁移为一个 24000 道尔顿的单峰，还原后的白细胞源性干扰素的残余活性也是如此。我们的数据表明，携带克隆 Hif-SN35-AH-L6 的大肠杆菌产生的干扰素在大小和跨物种活性方面与白细胞源性干扰素的一种分子类型相似。
