绒山羊早孕因子的活性检测及分离纯化  

Purification and characterization of early pregnancy factor from Liaoning Cashmere Goat serum

作　　者：张兴会[1] 宋先忱[1] 郭丹[1] 韩迪[1] 

机构地区：[1]辽宁省畜牧科学研究院,辽宁辽阳111000

出　　处：《现代畜牧兽医》2013年第11期8-12,共5页Modern Journal of Animal Husbandry and Veterinary Medicine

基　　金：高产绒量转基因绒山羊新品种培育(编号:2011ZX08008-002)

摘　　要：检测怀孕绒山羊早孕因子的活性，并对其进行分离纯化，以期为EPF快速检测方法的建立提供依据。采集妊娠1～4个月健康孕羊血清，通过DEAE-52离子交换层析、ConA—Sepharose4B亲和层析和Heparin SepharoseC16B亲和层析分离纯化技术，从孕羊血清中分离出纯化的EPF。用E玫瑰花环抑制试验对各阶段产物EPF活性进行检测。结果表明，受精后的绒山羊在22～30h能够检测出早孕因子活性，怀孕羊和未怀孕羊血清的E玫瑰花环抑制滴度差异显著（P〈0．05），纯化出的EPF通过E玫瑰花环抑制试验显示具有活性，其分子质量为47．7ku，等电点为6．7。The study was done to detect the activity of EPF and purify EPF from pregnant Cashmere Goat serum using rosette inhibition test. Serum was obtained from pregnant Cashmere Goat of 1-4 months. Rosette inhibition test was used to detect the activity o f EPF in pregnant Cashmere Goat, and then serum was purified by DEAE-Sepharose, Con A Sepharose 4B affinity chromatography and Hepar in Sepharose C1 6B affinity chromatography. The purified products were detected by SDS-PAGE and isoelectric points were detected. After insemination, the RIT has the potential to distinguish pregnant status of Cashmere Goat at 22-30 h. The result s indicated that RIT signifi-cantly deviated （P〈0. 05） between pregnant and non-pregnant Cashmere Goat. The EPF＇ s activity of Cashmere Goat was detected in the final purified products and it＇s molecular weight was 47.7 ku, the isoelectric point 6.7.

关 键 词：绒山羊 早孕因子 妊娠状态 孕血清 

分 类 号：S831[农业科学—畜牧学]