55K猪卵透明带抗原(ZP_3)的分离纯化  

ISOLATION AND PURIFICATION OF 55K PORCINE ZONA PELLUCIDA ANTIGEN (ZP_3)

作　　者：谢琪璇[1] 张江兰[1] 陈海玲[1] 潘善培[1] 

机构地区：[1]暨南大学生殖免疫研究中心

出　　处：《暨南大学学报（自然科学与医学版）》1992年第1期115-121,共7页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国家自然科学基金

摘　　要：为了获得足量的猪卵透明带55K组分(ZP_3),以便研究其在高等灵长类恒河猴的免疫抗生育效果、生化和免疫学特性,我们根据国内条件建立了一套从猪卵巢分离纯化透明带55K组分的方法,该法包括用排刀切割卵巢回收卵子,热溶提取卵透明带组分(SIZP),Sephacryl S-400柱层析和Hydroxylapatite(HTP)柱层析分离纯化ZP_3、ZP_3的复性等步骤。采用本法,获得了在SDS-PAGE图谱上呈单一蛋白染色带并保留其抗原性的ZP_3纯品。每2000个猪卵巢可得ZP_3纯品10mg以上。Studies were carried out to isolate and purify the 55K porcine zona pellucida antigen (ZP_3) from ovaries collected by veterinarians during castrating pigs for obtaining enough ZP_3 to study its immunocontrception effect, biochemical and immunological properties. A series of methods for isolating and purifying ZP_3 based on our lab's conditions were set up. Ocytes and zona pellucida were collected by cutting the surface of ovaries with a row of razor blades manually, washing ovaries with PBS which contains 2mM sodium citrate, and passing the suspension of oocytes, zona pellucida and follicle cells through 500, 300, 200,120. 75and 45 um nylon screens. lsolated oocytes and zona pellucida were abtained by washing 7.5 and 45 um screens. Solubilized zona pellucida (SIZP) were directly prepared hy heating the mixture of oocytes and zona pellucida in zona buffer at 75°(in water bath. ZP_3 was isolated and purified by Sephacryl S-400 and hydroxylapatite(HTP) column chromatography. By these methods, pure ZP_3 was obtained, which still retains the ZP_3 antigenicity and shows a single band pattern in SDS-PAGE. Over 10mg pure ZP_3 can be prepared from 2000 ovaries.

关 键 词：卵透明带 分离 纯化 ZP3 

分 类 号：Q492.5[生物学—生理学]