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.
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)
关键词
绒山羊
早孕因子
妊娠状态
孕血清
Cashmere Goat
Early pregnancy factor:Pregnant state
Pregnancy serum