长角血蜱磷酸丙糖异构酶的真核表达及免疫保护力分析  

Eukaryotic expression and immune protection analysis of triosephosphate isomerase in Haemaphysalis longicornis

作　　者：吴品兴 梁娜 董红猛 肖舒文 胡永红[1] WU Pinxing;LIANG Na;DONG Hongmeng;XIAO Shuwen;HU Yonghong(College of Life Sciences,Hebei Normal University,Shijiazhuang 050024,China)

机构地区：[1]河北师范大学生命科学学院,河北石家庄050024

出　　处：《中国兽医学报》2021年第9期1774-1779,共6页Chinese Journal of Veterinary Science

基　　金：教育部春晖计划合作科研资助项目(Z2017-018);河北师范大学重点基金资助项目(L2020Z06)。

摘　　要：在前期克隆长角血蜱磷酸丙糖异构酶TIM基因基础上,将密码子优化后的TIM基因合成于真核质粒pPICZaA中,并转入毕赤酵母X33进行真核表达。经镍柱纯化得到重组蛋白rTIM,相对分子质量大小约28 kDa。Western blot结果表明rTIM具有免疫原性。将rTIM与佐剂混合免疫新西兰大白兔,ELISA结果显示,免疫rTIM的新西兰大白兔与对照组相比具有较高的抗体水平。蜱叮咬试验结果显示,与对照组相比,试验组雌蜱饱血体质量(163.30±1.40)mg、产卵量(52.23±1.41)mg和卵孵化率(29.26±2.43)%显著降低,分别减少11.02%,16.00%和44.69%;计算得出rTIM免疫保护率为58.40%。研究结果为进一步筛选抗蜱疫苗提供了理论依据。Based on the cloning of the triosephosphate isomerase gene from Haemaphysalis longicornis,the codon-optimized TIM gene was synthesized and cloned into the eukaryotic plasmid pPICZaA,then it was transformed to Pichia pastoris X33 for eukaryotic expression.The recombinant protein rTIM was purified by nickel column.The result of SDS-PAGE showed that its molecular weight was about 28 kDa.Western blot results showed that the recombinant protein rTIM has immunogenicity.New Zealand white rabbits were immunized with a mixture of rTIM and adjuvant.ELISA results showed that rabbits immunized with rTIM had higher antibody levels than that the control group.The results of the tick bite experiment showed that compared with the control group,engorgement weight(163.30±1.40)mg,ovipositions(52.23±1.41)mg and hatchability(29.26±2.43)%of female ticks in the experimental group were significantly reduced by 11.02%,16.00%and 44.69%respectively.The immune protection rate of rTIM was 58.40%.The study provides a theoretical basis for further screening of anti-tick vaccines.

关 键 词：长角血蜱 磷酸丙糖异构酶 真核表达 抗蜱疫苗 

分 类 号：S852.746[农业科学—基础兽医学]