中文摘要：

目的构建pET28a-FAAP-H is原核表达载体,表达H is-FAAP融合蛋白,制备FAAP蛋白的多克隆抗体。方法构建pET28a-FAAP-H is原核表达载体,转化到大肠杆菌BL21中并诱导表达H is-FAAP融合蛋白,用纯化的FAAP蛋白免疫昆明小白鼠后,制备多克隆抗体,通过W estern blotting检验抗血清的特异性和灵敏性。结果成功构建pET28a-FAAP-H is原核表达载体。在大肠杆菌BL21中经1 m M的异丙基-β-D-硫代半乳糖苷（isopropy-β-D-thiogalactoside,IPTG）37℃诱导4 h后,融合蛋白有很高水平的表达。W esternblotting结果显示FAAP抗体能够有效地检测人血管内皮脐静脉细胞内FAAP蛋白的表达。结论成功地对FAAP进行了原核表达、纯化,抗FAAP小鼠的多克隆抗体具有较好的特异性,为进一步研究FAAP的结构与功能奠定了坚实的基础。

英文摘要：

【Objective】 This study aims to construct a His-tagged prokaryotic expression vector named pET28a-FAAP-His and express it in Escherichia coli to obtain anti-FAAP polyclonal antibody.【Methods】 A His-tagged prokaryotic expression vector pET28a-FAAP-His was constructed and transfected into E.coli BL21.The recombinant FAAP protein was resolved by renaturation of gradient urea dialysis after purification,and then it was used as antigen to immune mice to obtain polyclonal antibody.The specificity and the antiserum sensitivity of the mouse anti-FAAP antibody were assayed and confirmed by Western blot.【Results】 The results showed that the vector pET28a-FAAP-His was successfully constructed,The FAAP protein was highly expressed while treated with1mM IPTG at 37°C for 4 hours in E.coli BL21,thus the optimized protein expression induction condition was confirmed.Western blot showed that anti-FAAP polyclonal antibody can be used to detect the expression of FAAP protein in human umbilical vein endothelioid cells.【Conclusion】 We successfully constructed the His-tagged prokaryotic expression vector pET28a-FAAP-His and obtain anti-FAAP polyclonal antibody,which is highly specific and sensitive.The work has laid a solid foundation for further study to the structure and function of FAAP protein.