红螯光壳螯虾胚胎及仔虾发育过程中卵黄磷蛋白的ELISA检测

建立了间接竞争酶联免疫吸附反应(ELISA)方法,对红螯光壳螯虾(Cherax quadricarinatus)胚胎及仔虾发育过程中的卵黄磷蛋白含量及其亚基组分进行了研究。该方法对卵黄磷蛋白具有良好的特异性,有效检测范围为31.25~250 ng/ml。结果表明,在发育初期胚胎先降解分子量比卵黄磷蛋白大的蛋白;亚基中,分子量较大和较小的亚基都先被消耗;胚胎内卵黄磷蛋白含量总体上呈下降趋势,其中在卵裂囊胚阶段后略有上升(3.19%),至后无节幼体期卵黄磷蛋白含量达最高(4.67%),之后不断下降,到仔虾离开母体独立生活时,含量只剩下卵裂期的1/4。

Detection of Vitellin Concentration Changes during Embryonic and Laval Development of Redclaw Crayfish,Cherax quadricarinatus,by ELISA

An indirect competitive enzyme linked immunosorbent assay(ELISA) for detecting vitellin in embryonic and laval Redclaw Crayfish,Cherax quadricarinatus was developed.This method was sensitive to the vitellin,with the effective range of 31.25-250 ng/ml.The results indicated that,during embryonic development,proteins with a molecular weight larger than that of vitellin were degraded,and both large and small molecular weight protein subunits were consumed first.The concentration of vitellin decreased during development.Its concentration was a little increased at cleavage stage and blastula stage(3.19%),reached the highest level in nauplius stage(4.67%),and then decreased remarkably when the laval could live independently,at which time the concentration of the vitellin was only 1/4 of that in fertilized eggs.