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为了从海南单柄蛛Ornithoctonus hainana毒素中获得对哺乳动物无毒而对昆虫有毒杀作用的单一蛋白组分,应用反相层析和阳离子交换对海南单柄蛛粗毒素进行分离纯化。分别通过胸腔注射、腹腔注射和饲喂的方法测定各毒素组分对蟋蟀、小鼠和棉铃虫的毒性作用,并采用改进的寇式法计算LD50。分离得到一种新的杀虫肽,命名为海南单柄蛛毒素a(Hainana toxica,HNTXa),聚丙烯酰胺等点聚焦电泳(IEF-PAGE)分析其等电点为8.15～8.45。杀虫肽HNTXa对棉铃虫和蟋蟀有较强的抑制作用。棉铃虫经饲喂浸沾HNTXa的棉花叶片72h后,死亡率超过30%;HNTXa对蟋蟀的LD50值为58μg/g,LD50的95%置信区间为55～62μg/g;HNTXa对哺乳动物小鼠无明显毒性。
In order to obtain a single protein component insensitive to insects from the Ornithoctonus hainana toxin in Hainan, the monoterper venom of Hainan was isolated and purified by reverse phase chromatography and cation exchange. The toxic effects of each toxin component on crickets, mice and cotton bollworm were determined by intrathoracic injection, intraperitoneal injection and feeding, respectively. LD50 was calculated by the improved Kaplan method. A new insecticidal peptide was isolated and named as Hainana toxica (HNTXa) and polyacrylamide. The isoelectric point was 8.15 ~ 8.45 by IEF-PAGE. The insecticidal peptide HNTXa has a strong inhibitory effect on cotton bollworm and crickets. The mortality of cotton bollworm was over 30% after 72h exposure to HNTXa-infected cotton leaves. The LD50 of HNTXa to crickets was 58μg / g, and the 95% confidence interval of LD50 was 55 ~ 62μg / g. No obvious toxicity.