College of Animal Science, Xinjiang Agricultural University, Urumqi 830053, Xinjiang, China; Parasitology Laboratory, College of Veterinary, Xinjiang Agricultural University, Urumqi 830053, Xinjiang, China.
Parasitology Laboratory, College of Veterinary, Xinjiang Agricultural University, Urumqi 830053, Xinjiang, China.
Vet Parasitol. 2020 Mar;279:109043. doi: 10.1016/j.vetpar.2020.109043. Epub 2020 Jan 30.
Dermacentor marginatus is one of the main tick species in northwestern China, and is a vector of various tick-borne pathogens. Tick control method largely depends on chemical agents, but the disadvantages of using such approach would cause environmental damage and the risk of developing tick resistance to acaricides. Vaccination of tick protective antigen is an eco-friendly approach which is an alternative and promising method to mitigate tick infestation in livestock. In the study, a mu-class glutathione S-transferase (GST) sequence of D. marginatus was cloned and the recombinant protein (rDmGST) was expressed. Transcriptional level of the GST was measured together with native GST activity of the tick. Finally, A vaccine trial on rabbits against D. marginatus was proceeded to evaluate the anti-tick effect of rDmGST. Results reveled that the CDs of the D. margiantus glutathione S-transferase mu 1 gene has 669 base pair nucleotide sequence encoding a 223 amino acid. The deduced GST protein sequence had over 95 % similarity with that of D. variabilis. The rDmGST was efficiently expressed soluble and purified by His trap affinity chromatography. Enzyme activity of native GST and transcriptional profiles of the GST showed up-regulation in different stages and organs of D. marginaus during blood feeding. Polyclonal antibody reacted with rDmGST in Western blotting. Tick challenge on rDmGST inoculated rabbits showed reductions in adult female engorgement rate, total egg mass and egg hatching rate with an overall vaccine efficacy of 43.69 %. The results of the experiment indicated the GST has potential value to be an effective protective antigen of D. marginatus.
边缘革蜱是中国西北地区的主要蜱种之一,是多种蜱传病原体的载体。蜱的控制方法主要依赖于化学药剂,但这种方法的缺点会造成环境破坏和蜱对杀蜱剂产生抗药性的风险。蜱保护性抗原的疫苗接种是一种环保的方法,是减轻牲畜蜱虫感染的替代和有前途的方法。本研究中,克隆了边缘革蜱的一个 mu 类谷胱甘肽 S-转移酶(GST)序列,并表达了重组蛋白(rDmGST)。同时测量了 GST 的转录水平和蜱的天然 GST 活性。最后,对兔子进行了针对 D. marginatus 的疫苗试验,以评估 rDmGST 的抗蜱效果。结果表明,边缘革蜱谷胱甘肽 S-转移酶 mu 1 基因的 CDs 具有 669 个碱基对的核苷酸序列,编码 223 个氨基酸。推导的 GST 蛋白序列与 D. variabilis 的相似性超过 95%。rDmGST 通过 His 亲和层析可有效表达可溶性和纯化。天然 GST 的酶活性和 GST 的转录谱显示,在边缘革蜱吸血的不同阶段和器官中均有上调。多克隆抗体在 Western blot 中与 rDmGST 反应。rDmGST 接种的兔子受到蜱虫的挑战后,雌性成虫的饱血率、总产卵量和卵孵化率均降低,总疫苗效力为 43.69%。实验结果表明,GST 具有作为边缘革蜱有效保护性抗原的潜力。