【摘要】 用蒸馏水沉淀法提取大草蛉(ChrysopaseptempunctataWesmael)的卵黄蛋白,免疫家兔,并将所得抗血清经雄虫匀浆液吸收,获得抗卵黄蛋白的特异抗体,经纯化后,用辣根过氧化物酶标记抗体。采用酶联免疫测定(ELISA)直接法,系统测定了大草蛉成虫期脂肪体、血淋巴和卵巢中卵黄蛋白的动态变化。结果表明:脂肪体是卵黄原蛋白合成的场所,卵黄原蛋白的合成始于羽化后第4d;脂肪体、血淋巴中卵黄原蛋白的滴度在羽化后第4d开始迅速上升,至成虫期的第7d达到高峰期;在羽化后第4d的卵巢中可以检测到卵黄蛋白,此后其含量迅速上升,到成虫期的第7d达到高峰期。经SDS-PAGE分析,结果表明:大草蛉的卵黄蛋白由165kD和46kD的大小2个亚基组成。更多还原

【Abstract】 The paper is the establishment of an ELISA for measurement of vitellin in Chrysopa septempunctata. The vitellin of Chrysopa septempunctata was extracted with cold distilled water. A polyclonal antibody against vitellin was prepared in male rabbits and then it was made the specific antibody for vitellin by absorbtion with male worm homogenate. As the result, specific antibody could be obtained which was anti-vitellin. Through purification, HRP was used to mark antibody, then ELISA was established to detect vitellin content in the Chrysopa septempunctata fat body, hemolymph and ovary. The result implied that the vitellogenin was synthesized in fat body at the beginning of the fourth day after eclosion. The content of vitellogenin in fat body and haemolymph increased on the fourth day after eclosion quickly and reached the peak stage on the seventh day in adult period. The vitellin in ovary could be detected on the fourth day after eclosion. Then the content of vitellin increased quickly and reached the peak stage on the seventh day in adult period. SDS-PAGE analysis showed that the vitellin was composed of two subunits, one large 165 kD and one small 46 kD.更多还原