水产学报2011，Vol.35Issue(4)：614-619,6.DOI:10.3724/SP.J.1231.2011.17255
牙鲆皮肤黏液免疫球蛋白的纯化及部分特性分析
Purification and characterizations of skin mucus immunoglobulin in Japanese flounder (Paralichthys olivaceus)
摘要
Abstract
Teleost fish have a unique physical barrier composed of mucus and skin which act as the first line of defence.It has been shown that lymphocytes are present in the epidermis of fish, and the cutaneous mucus of several fish species contains immunoglobulin (Ig).As fish mucosal composition was complicated and the content of Ig was low, it is difficult to obtain high purity mucus Ig.In this study, skin mucus Ig of Japanese flounder (Paralichthys olivaceus )was purified by the combination of saturated ammonium sulfate precipitation, Sephacryl S-300 gel filtration chromatography and DEAE Sepharose chromatography, and partial characteristics of purified proteins were analyzed by SDS-PAGE and Western-blot.According to SDSPAGE,many other proteins were removed from mucus by precipitation of 30％ and 50％ saturated ammonium sulfate solution, and Ig that was crudely extracted could reach a high purity by further purification using Sephacryl S-300.After being finally purified by DEAE Sepharose chromatography, skin mucus Ig has only two bands of 72 and 26 ku,initially presumed as the heavy chain and light chain of Japanese flounder mucus Ig.Western-blot analysis showed that the monoclonal antibody produced from serum Ig could recognize the heavy chain of mucus Ig(72 ku).关键词
牙鲆/肤黏液免疫球蛋白/盐析/凝胶过滤层析/离子交换层析Key words
Paralichthys olivaceus/ mucus immunoglobulin/ ammonium sulfate precipitation/ gel filtration chromatography/ DEAE Sepharose chromatography分类
生物科学引用本文复制引用
许国晶,绳秀珍,唐小千,邢婧,战文斌..牙鲆皮肤黏液免疫球蛋白的纯化及部分特性分析[J].水产学报,2011,35(4):614-619,6.基金项目
国家自然科学基金项目(30771648,31072232) （30771648,31072232）
