激光生物学报, 2018, 27 (3): 260, 网络出版: 2018-09-07
利用DNA免疫技术制备ASB11蛋白的多克隆抗体
Preparation of Polyclonal Antibody Against ASB11 Protein by DNA Immunization
摘要
ASB11参与胚胎神经祖细胞的发育、再生性肌发生以及泛素化等过程,但其机制仍不清楚。为了进一步研究斑马鱼Asb11基因的作用机制,本研究采用DNA免疫技术制备了ASB11多克隆抗体;利用斑马鱼Asb11的cDNA构建pCAGGS-P7/ASB11重组表达质粒,肌肉注射入6-8周龄的BALB/c小鼠体内,诱导抗原蛋白的表达和免疫应答的发生。结果显示,制备的pCAGGS-P7/ASB11重组质粒具有较好的免疫原性;将提取的抗血清进行Western-blot和免疫荧光检测,显示所制备的多克隆抗体抗体效价为1∶400,抗血清抗体能特异的结合ASB11蛋白。本研究为后续的功能研究奠定了基础。
Abstract
ASB11 is involved in the development of embryonic neural progenitor cells, regenerative myogenesis and ubiquitination, but its detailed mechanism of action has not been known. In order to further analyze the mechanism of zebrafish Asb11 gene, the anti-ASB11 specific polyclonal antibody by immunizing mice with DNA was prepared. The eukaryotic expression vector pCAGGS-P7/ASB11 was constructed by using cDNA of zebrafish Asb11. BALB/c mice of 6-8 weeks old were immunized with pCAGGS-P7/ASB11 to prepare anti-serum against ASB11. The results showed that the pCAGGS-P7/ASB11 recombinant plasmid was constructed with a rather good immunogenicity. The antibody valence of anti-serum obtained from the immunized mice was 1∶400. Western blotting and indirect immunofluorescence showed that the anti-serum prepared by immunizing mice with DNA could specifically recognize the ASB11 protein. This research established afoundation to reveal the mechanism of ASB11 protein.
蔡英桂, 尹丽阳, 王宏波, 陈宇, 曾蓉, 吴秀山, 叶湘漓. 利用DNA免疫技术制备ASB11蛋白的多克隆抗体[J]. 激光生物学报, 2018, 27(3): 260. CAI Yinggui, YIN Liyang, WANG Hongbo, CHEN Yu, ZENG Rong, WU Xiushan, YE Xiangli. Preparation of Polyclonal Antibody Against ASB11 Protein by DNA Immunization[J]. Acta Laser Biology Sinica, 2018, 27(3): 260.