Yang Xiaolong, Yu Zhijun, He Yanjie, Xu Xiaoli, Gao Zhihua, Wang Hui, Chen Jie, Liu Jingze
Key Laboratory of Animal Physiology, Biochemistry and Molecular Biology of Hebei Province, College of Life Sciences, Hebei Normal University, Shijiazhuang, 050024, China.
Exp Appl Acarol. 2015 Mar;65(3):377-88. doi: 10.1007/s10493-014-9866-z. Epub 2014 Dec 19.
Vitellin (Vt) was purified from eggs of parthenogenetic bush tick Haemaphysalis longicornis by gel filtration and ion exchange chromatography. Our results revealed that only one single Vt existed in parthenogenetic bush tick, and the purified Vt was proved to be a hemoglycolipoprotein consisting of nine polypeptides with molecular weights of 203, 147, 126, 82, 74, 70, 61, 47 and 31 kDa, respectively. Polyclonal antibody and monoclonal antibody against Vt were produced using the purified Vt. The change in vitellogenin (Vg) and Vt levels over time of the parthenogenetic H. longicornis was established, and the Vg content in haemolymph and Vt in ovary at different feeding or engorgement statuses was also determined using a double antibody sandwich enzyme-linked immunosorbent assay. The Vg level in haemolymph was distinctly increased on the day of engorgement (1.785 mg/mL) and continued to increase until 2nd day post-engorgement (5.611 mg/mL). There was a slight decrease in Vg level after 4 days of engorgement, and a second peak was observed on day 2 post-oviposition (10.774 mg/mL). Subsequently, Vg content continuously decreased and reached a low level on the 10th day post-oviposition. The Vt content in ovary continuously increased once the female reached its critical weight (0.024 mg per female), and reached the maximum level on day 2 post-oviposition (1.942 mg per female). Afterwards, Vt content rapidly decreased.
通过凝胶过滤和离子交换色谱法从孤雌生殖的长角血蜱卵中纯化出卵黄磷蛋白（Vt）。我们的结果表明，孤雌生殖的长角血蜱中仅存在一种单一的Vt，并且纯化后的Vt被证明是一种由9种多肽组成的血糖脂蛋白，其分子量分别为203、147、126、82、74、70、61、47和31 kDa。使用纯化后的Vt制备了针对Vt的多克隆抗体和单克隆抗体。确定了孤雌生殖的长角血蜱中卵黄原蛋白（Vg）和Vt水平随时间的变化，并使用双抗体夹心酶联免疫吸附测定法测定了不同取食或饱血状态下血淋巴中的Vg含量和卵巢中的Vt含量。饱血当天血淋巴中的Vg水平明显升高（1.785 mg/mL），并持续升高直至饱血后第2天（5.611 mg/mL）。饱血4天后Vg水平略有下降，产卵后第2天观察到第二个峰值（10.774 mg/mL）。随后，Vg含量持续下降，并在产卵后第10天降至低水平。一旦雌蜱达到临界体重（每只雌蜱0.024 mg），卵巢中的Vt含量就会持续增加，并在产卵后第2天达到最高水平（每只雌蜱1.942 mg）。此后，Vt含量迅速下降。
