Kashima T, Iwata H, Inoue T
Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University, Japan.
J Vet Med Sci. 1999 Jun;61(6):705-7. doi: 10.1292/jvms.61.705.
Bovine interleukin-2 (bIL-2) was expressed in insect cells infected with recombinant baculovirus. The cDNA for bIL-2 was amplified by reverse transcriptional polymerase chain reaction and recombinant baculovirus was constructed by homologous recombination. The recombinant baculovirus was plaque-purified, amplified, and then infected to Sf9 cells for expression of recombinant bIL-2 (rbIL-2). In result, the protein band corresponding to rbIL-2 could be detected on SDS-PAGE with coomassie brilliant blue staining and on immunoblot analysis reacted with mouse antiserum against bIL-2. In addition, cell proliferation assay for bIL-2 activity demonstrated that the culture supernatant of Sf9 cells infected with recombinant baculovirus enhanced the proliferation of bovine peripheral blood mononuclear cells in a dose-dependent manner.
牛白细胞介素-2(bIL-2)在感染重组杆状病毒的昆虫细胞中表达。通过逆转录聚合酶链反应扩增bIL-2的cDNA,并通过同源重组构建重组杆状病毒。对重组杆状病毒进行噬斑纯化、扩增,然后感染Sf9细胞以表达重组bIL-2(rbIL-2)。结果,在考马斯亮蓝染色的SDS-PAGE上可检测到与rbIL-2对应的蛋白条带,并且在免疫印迹分析中与抗bIL-2的小鼠抗血清发生反应。此外,bIL-2活性的细胞增殖试验表明,感染重组杆状病毒的Sf9细胞培养上清液以剂量依赖性方式增强了牛外周血单个核细胞的增殖。