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c DNA构建p CAGGS-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p CAGGS-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。
ASBll is involved in the development of embryonic neural progenitor cells, regenerative myogenesis and ubiquitination, but its detailed mechanism of aetion has not been known. In order to further analyze the mechanism of zebrafish Asbl 1 gene, the anti-ASB11 specific polyelonal antibody by immunizing mice with DNA was prepared. The eu- karyotie expression vector pCAGGS-P7/ASB11 was constructed by using eDNA of zebrafish Asbl 1. BALB/e mice of 6-8 weeks old were inmmnized with pCAGGS-P7/ASBI 1 to prepare anti-serum against ASB11. The results showed that the pCAGGS-P7/ASB11 recombinant plasmid was constructed with a rather good inmmnogenieity. The antibody valence of anti-serum obtained from the inmmnized mice was 1 ： 400. Western blotting and indirect immunofluoreseenee showed that the anti-serum prepared by immunizing mice with DNA could specifically recognize the ASB11 protein. This research es- tablished afoundation to reveal the mechanism of ASB11 protein.
作者
蔡英桂
尹丽阳
王宏波
陈宇
曾蓉
吴秀山
叶湘漓
CAI Yinggui;YIN Liyang;WANG Hongbo;CHEN Yu;ZENG Rong;WU Xiushan;YE Xiangli(College of Medicine,Hunan Normal University,Changsha 410013,Hunan,China;College of Life Sciences,Hunan Normal University,Changsha 410081,Hunan,China;The Second XiangYa Hospital of Central South University,Changsha 410011,Hunan,China)
出处
《激光生物学报》
CAS
2018年第3期260-264,280,共6页
Acta Laser Biology Sinica
基金
国家自然科学基金面上项目(81270291)
湖南师范大学科研基金博士科研启动项目
湖南省大学生研究性学习和创新性实验计划项目
