Interferon-stimulated gene 15 (ISG15) is known to participate in innate and adaptive immune responses against viral infections. Classical swine fever virus (CSFV) causes a global, economically important and highly contagious disease in pigs. To explore theinteraction between ISG15 and CSFV, the ISG15gene was amplified, cloned and expressed in E. coliBL21 cells. The expressed ISG15 protein was purified by Ni-NTA agarose column and used to prepare a polyclonal mouse antibody. The anti-ISG15 antibody produced in mice could react with ISG15 expressed in the prokaryotic BL-21 cells as well as natural swine ISG15 expressed in PK-15 cells. This polyclonal antibody against swine ISG15 was used to perform immunofluorescence assay, western blot and flow cytometryfor the detection of the ISG15 protein and to identify that ISG15 was up-regulatedin PK-15 cells after infection with CSFV. This polyclonal antibody against ISG15 protein is importantfor further research on the interaction between CSFV and ISG15.
Faculty of Veterinary Science, Chulalongkorn University
Cai, Xin-Bin; Hou, Hua-Lin; Zhang, Li-Yuan; Tang, Rong-Ze; Yin, Shan; Li, Xiao-Quan; Zhang, Ke; Zhao, Qian-Nan; Ying, Xue; Su, Li-Juan; Sun, Shi-Kai; Li, Xiao-Ning; and Luo, Ting Rong
"Preparation of swine ISG15 polyclonal antibody and its application in classical swine fever virus,"
The Thai Journal of Veterinary Medicine: Vol. 50:
3, Article 15.
Available at: https://digital.car.chula.ac.th/tjvm/vol50/iss3/15