趋化因子CCL28基因的克隆表达及其多抗制备  

Cloning and expression of chemokine CCL28 gene and its polyclonal antibody preparation

作　　者：陆文静 王礼学 王国相 王伟康 谢泉[1] 谢菁 邵红霞[1] 高巍[1] 秦爱建[1] 叶建强[1] LU Wenjing;WANG Lixue;WANG Guoxiang;WANG Weikang;XIE Quan;XIE Jing;SHAO Hongxia;GAO Wei;QIN Aijian;YE Jianqiang(Key Laboratory of Poultry Preventive Medicine,Ministry of Education/Jiangsu Key Laboratory of Animal Preventive Medicine/Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Diseases and Zoonoses/Agricultural Science and Technology Development Research Institute/Joint Laboratory of International Cooperation in Agriculture and Agricultural Product Safety,Ministry of Education/College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Department of Radiation Oncology Nanjing Hospital Affiliated (Nanjing Second Hospital),Nanjing University of Traditional Chinese Medicine,Nanjing 210003,China;Nanjing Zhitai Biomedical Technology Company Limited,Nanjing 210061,China)

机构地区：[1]扬州大学兽医学院/禽类预防医学教育部重点实验室/江苏省动物预防医学重点实验室/江苏省动物重要疫病与人兽共患病防控协同创新中心/农业科技发展研究院/教育部农业与农产品安全国际合作联合实验室,江苏扬州225009 [2]南京中医药大学附属南京医院(南京市第二医院)放疗科,南京210003 [3]南京至泰生物医药科技有限公司,南京210061

出　　处：《扬州大学学报（农业与生命科学版）》2019年第4期44-48,54,共6页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：江苏高校优势学科建设工程项目(PAPD)

摘　　要：将原核表达质粒pGEX-6P-1-CCL28转化至大肠埃希菌BL21中,经IPTG诱导,获得了GST-CCL28可溶性融合表达产物。利用纯化的GST-CCL28蛋白免疫BALB/c小鼠制备了抗GST-CCL28的多克隆抗体。同时将CCL28基因亚克隆进真核表达载体,经测序验证后命名为pcDNA3.1-CCL28。间接免疫荧光及Western blot试验结果进一步证明,所获得的抗GST-CCL28多克隆抗体能特异性的识别转染pcDNA3.1-CCL28质粒的DF-1细胞内高表达的CCL28蛋白。这一研究为制备抗CCL28单克隆抗体和探究与CCL28相关的生物学功能及其机制奠定了良好的基础。In this study, the prokaryotic expression plasmid pGEX-6 P-1-CCL28 was transformed into BL21 and the soluble fusion expression of GST-CCL28 induced by IPTG was obtained. Moreover, the polyclonal antibody against GST-CCL28 was prepared by immunizing BALB/c mice with the purified GST-CCL28 protein. The chemokine CCL28 gene was also subcloned into eukaryotic expression vector, named as pcDNA3.1-CCL28 after confirmed by sequencing. Indirect immunofluorescence and Western blot assay revealed that the polyclonal antibody against GST-CCL28 generated could specifically recognize the CCL28 protein expressed in DF-1 cells transfected with pcDNA3.1-CCL28. All these data laid a solid foundation for the preparation of monoclonal antibodies against CCL28 and for the exploration of biological functions of CCL28.

关 键 词：趋化因子CCL28 克隆表达 多克隆抗体 

分 类 号：Q939.91[生物学—微生物学]