猪IL-22的原核表达及其活性研究  

Prokaryotic expression and bioactivity of porcine interleukin-22

作　　者：应兰[1,2] 薛美[2] 李麟[2] 符芳[2] 冯力[2] 赵静[1] 刘平黄[2] 

机构地区：[1]青海大学农牧学院,青海西宁810003 [2]中国农业科学院哈尔滨兽医研究所,黑龙江哈尔滨150001

出　　处：《中国预防兽医学报》2016年第6期489-492,共4页Chinese Journal of Preventive Veterinary Medicine

基　　金：中央级公益性科研院所基本科研业务费专项(0302014005)

摘　　要：为表达猪白细胞介素22(PoIL-22)并研究其活性,本研究从猪小肠上皮细胞IPEC-J2中扩增出568 bp的PoIL-22编码序列,并以其为模板进一步扩增去除信号肽的PoIL-22 471 bp序列。将该去信号肽基因克隆到原核表达载体pET-30a(+)中,转化至大肠杆菌BL21(DE3)中诱导表达了重组蛋白PoIL-22(rPoIL-22),经过Ni-NTA亲和层析纯化。此外,以纯化的rPoIL-22刺激IPEC-J2后,能够上调IPEC-J2表达猪beta防御素2,并表现剂量依赖效应。由此证明本实验中表达的rPoIL-22具有生物学活性。rPoIL-22的制备为进一步研究其在猪粘膜免疫中的作用奠定了基础。Interleukin-22, a member of IL-10 super family cytokines, is primarily secreted by T lymphocytes and innate lymphoid cells. Recently growing studies from mouse or human IL-22 demonstrate that IL-22 primarily target mucosal epithelial cells, play a vital role in the repair of mucosal wound, inflammatory response, and fight against infection. However, there is no report of porcine IL-22 （PolL-22） until now. In this study, we amplified IL-22 from swine intestinal epithelia IPEC-J2 cells by RT-PCR, and cloned into prokaryotic pET-30a for expressing in E.coli. The expressed recombinant porcine IL-22（rPolL-22） was purified through Ni-NTA affinity chromatography. The beta-defensin-2（BD-2） expression was up-regulated in IPEC-J2 after being treated by rPolL-22, and exhibited in a dose-dependent manner. This demonstrates that PolL-22 has bioactivities in innate immunity, which provides a basis for further studying the role of PolL-22 in mucosat immunity.

关 键 词：猪IL-22 蛋白表达 猪小肠上皮细胞 黏膜免疫 

分 类 号：S852.4[农业科学—基础兽医学]