目的 构建北方羽螨pET28a-CatD载体，为表达北方羽螨的组织蛋白

Objective To construct the pET28a-CatD vector of northern feather mite, and provide a scientific basis for the expression of cathepsin D of northern feather mite and whether the antigen protein can be used as a vaccine against chicken northern mite. <br>Methods The mites on chickens in Hainan were collected, and the morphology of the northern feather mites was preliminarily identified by microscopic observation. The genomic DNA of northern feather mites was extracted. Using this as a template, primers were designed, and then CatD was amplified by PCR. Then it was constructed on the pMD18-T cloning vector, and then transformed into the DH5α cloning strain, the plasmid was extracted, identified by enzyme digestion, and finally CatD was recovered and connected to pET28a. <br>Results The CatD gene was successfully obtained and constructed into the pMD18-T vector and the prokaryotic expression plasmid pET28a vector. Finally, pET28a-CatD was successfully obtained. <br>Conclusion The pET28a-CatD vector of northern feather mite was successfully constructed. <br><br>Key words Northern feather mite; carrier; construction

目的 构建北方羽螨pET28a-CatD载体，为表达北方羽螨的组织蛋白酶D以及研究该抗原蛋白是否能作为抗鸡北方羽螨的疫苗应用提供科学依据。<br>方法 采集海南地区鸡身上螨，通过显微镜观察形态初步鉴定为北方羽螨，提取出北方羽螨的基因组DNA，以其为模板，设计出引物，再用PCR扩增CatD，测序确定北方羽螨，然后构建到pMD18-T克隆载体上，再转化到DH5α克隆菌株，提质粒，经酶切鉴定，最后回收CatD连接到pET28a。<br>结果 成功获取到CatD基因，并将这个基因构建到pMD18-T载体和原核表达质粒 pET28a 载体上，最后成功获得pET28a-CatD。<br>结论 成功构建北方羽螨pET28a-CatD载体。<br><br>关键词 北方羽螨；载体；构建

Objective to construct pET28a CATD vector of northern feather mite, and to provide scientific basis for the expression of cathepsin D of northern feather mite and the application of the antigen protein as vaccine against northern feather mite.<br>method In this study, we collected the chicken mites from Hainan Province, and identified them as northern feather mites by microscopic observation. The genomic DNA of northern feather mites was extracted, which was used as template, designed primers, amplified CATD by PCR, sequenced northern feather mites, then constructed into pmd18-t clone vector, transformed into DH5 α clone strain, improved the quality particles, identified by enzyme cutting, and finally recovered CATD and connected to pET28a.<br>Results CATD gene was successfully obtained and constructed on pmd18-t vector and prokaryotic expression vector pET28a. Finally, pET28a CATD was successfully obtained.<br>Conclusion pET28a CATD vector was successfully constructed.<br>Keywords northern feather mite; carrier; construction<br>