【摘要】 为研究虾夷扇贝C型凝集素的母源传递及其抑菌作用,实验运用qRT-PCR技术检测了经鳗弧菌刺激后虾夷扇贝卵巢中C型凝集素的表达模式,比较分析了C型凝集素基因在正常虾夷扇贝和鳗弧菌刺激的虾夷扇贝所产的卵及其胚胎发育前期的存在与变化;通过抑菌实验研究了卵胞浆中C型凝集素抑制细菌存活的作用。结果表明,鳗弧菌刺激能够诱导虾夷扇贝卵巢中的C型凝集素mRNA表达量显著变化,最高表达量出现在刺激8 h后,为对照组的6.2倍;母体中C型凝集素可以传递给卵和胚胎,刺激组表达量极显著高于正常组,且表达量都随胚胎发育逐渐降低,至受精36 h时分别为正常对照卵的0.3和0.2倍;蛋白终浓度为200和400μg/mL的卵无细胞体系都具有一定的抑菌作用,与C型凝集素家族抗体反应后,细菌存活率显著上升,说明母源C型凝集素在抑制细菌存活方面发挥了重要的作用。更多还原

【Abstract】 Maternal transfer of immune factors from mother to eggs has been reported in many aquatic species,and these factors played important immune roles in developing embryos and larvae.However,the study on maternal immunity of shellfish remains lacking.In this study,maternal transfer and bacteriostasis of a C-type lectin in scallop(Patinopecten yessoensis)were analyzed.We detected the mRNA expression pattern of the C-type lectin gene in ovary and early developing larvae using the real-time quantitative PCR method,and tested the antibacterial activity of maternal C-type lectin in eggs by colony-forming unit(CFU)assay.The results of qRT-PCR showed that the expression of C-type lectin gene in ovary after bacterial challenge was time-dependent,and it was up-regulated gradually from 2 h after injection of bacteria and reached the peak at 8 h which was 6.2 times compared to the control group,and then dropped progressively to the original level.On the other hand,the fact that mRNA was found both in the eggs and larvae indicated that it could be transferred from mother to offspring.The mRNA expression in bacterial challenge group was significantly higher than those in normal group except for the 36 h point.The CFU assay revealed that the cytoplasm of eggs with protein concentration of 200 μg/mL and 400 μg/mL had lethal effect on the bacteria Vibrio anguillarum,and the function weakened significantly once the antibody of C-type lectin was added,which indicated the C-type lectin from mother had antibacterial activity.It is concluded that the maternal C-type lectin can transfer from mother to the offspring and has antibacterial activity.更多还原