| 摘要: |
| 本文报道了改进的柔嫩艾美耳球虫各阶段虫体纯化方法。用胃蛋白酶消化盲肠道组织，铜筛过滤除去大颗粒杂质后，以1．1mol·mL－1蔗糖离心漂浮卵囊，然后用小于0．5mol·mL－1蔗糖溶液反复漂洗，最后以5％次氯酸钠消毒处理，获得了大量纯化柔嫩艾美耳球虫卵囊。孢子化卵囊经研磨，消化，游离出子孢子后，过DEAE-纤维素52层析柱，以0．05mol·mL-1，Tris－HCl，（pH＝80，0．15mol·mL1-NaCl）洗脱，层析往高5cm，洗脱液水柱高3cm，流速为2mL·min－1，纯化出了子孢子。取人工感染球虫鸡的120h后的盲肠，游离出第二代裂殖子，铜筛过滤后，过柱方法同前，纯化出了第二代裂殖子。结果证明本法分离的子孢子和第二代裂殖子活性好，回收率达87％。 |
| 关键词: 柔嫩艾美耳球虫 卵囊 子孢子 第二代裂殖子 |
| DOI： |
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| 基金项目:国家攀登计划B类项目!85－44－01 |
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| A Improved Method for Purification Eimeria tenella Oocysts, Sporozoites and Second Generation Merozoites |
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| Abstract: |
| The present paper reported the improved methods for purification of Eimera tenella oocysts, sporozoites and second generation merozoites. The ceca containing oocysts was disrupted in disruptor and digested with pepsin; after filtration with different size filters, the primarily purified oocyst fraction was isolated by flotation in 1. 1mol .ml-1 sucrose and this crude material further purified by flotation repeatedly in <0. 5 mol' mL-1 sucrose; finally, the oocysts were strilized by incubating in 5 % cold NaOCl for 10min. After prepared crude material, the sporozoites and second generation merozoites were further purified by DEAE-cellulose 52 anion exchange chromatography with 0. 05 mol .mL-1 Tris-HCl pH = 8. 0 buffer, containing 0. 15 mol .mL-1 NaCl. With this method, the sporozoites and merozoites were still alive, and the mean recovery rate of sporozoites and merozoites was about 87 %. |
| Key words: DEAE-cellulose 52 anion exchange chromatography,Eimera tenells,oocysts,sporozoites,merozoites |
