NLRX1（NLR family member X1）是一种调控天然免疫应答的重要模式识别受体。首先应用PCR方法分别对鸡NLRX1基因的编码区和N端1～492 bp截短序列进行PCR扩增并克隆到原核表达载体pET-32a（+），以IPTG诱导重组菌表达，获得了重组NLRX1蛋白和N端截短表达蛋白（rNLRX1-part）。其次将纯化的截短重组蛋白rNLRX1-part免疫新西兰兔，以间接酶联免疫吸附实验（ELISA）检测制备的抗体效价为1∶512 000。应用Western blot方法分析显示，该抗体不仅可特异性结合原核表达NLRX1蛋白也可有效识别鸡DF-1细胞中过表达NLRX1蛋白。
NLR family member X1 (NLRX1) is an important pattern recognition receptor that regulates innate immune response. In this study, coding region sequence and 1-492 bp N-terminal truncated sequence of chicken NLRX1 gene were firstly cloned by PCR and inserted into prokaryotic expression pET-32a (+) vector. Expression of the recombinant NLRX1 protein and N-terminal truncated protein (rNLRX1-part) were induced by isopropyl β-D-Thiogalactoside (IPTG). Moreover, the recombinant rNLRX1-part protein was purified and immunized a New Zealand rabbit. The titer of anti-NLRX1 polyclonal antibody prepared by indirect enzyme-linked immunosorbent assay (ELISA) was 1∶512 000. The Western-blotting analysis showed the prepared antibody could not only specifically react with NLRX1 protein from prokaryotic expression but can effectively detect NLRX1 over-expression in chicken DF-1 cells.