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为揭示蝶蛹金小蜂Pteromalus puparum抑制寄主菜粉蝶Pieris rapae血细胞免疫反应的分子机制,克隆了菜粉蝶肌动蛋白、肌动蛋白解聚因子及微管蛋白cDNA,并研究了蝶蛹金小蜂寄生对其转录水平的影响。结果显示:菜粉蝶肌动蛋白cDNA ORF为1131bp,编码377aa,预测分子量为41.78kDa,pI为5.29,与其他昆虫肌动蛋白的相似性非常高,在90%以上。氨基酸组成特点和系统进化分析表明,克隆到的菜粉蝶肌动蛋白基因属细胞质型肌动蛋白。菜粉蝶肌动蛋白解聚因子cDNA全长为1243bp,ORF为447bp,编码149aa,预测分子量为16.97kDa,pI为7.11,与家蚕Bombyx mori、赤拟谷盗Tribolium castaneum、意大利蜜蜂Apis mellifera和桑粉介壳虫Maconellicoccus hirsutus肌动蛋白解聚因子的相似性分别为97%,87%,89%和72%。菜粉蝶微管蛋白cDNA全长为1757bp,ORF为1344bp,编码448aa,预测分子量为50.38kDa,pI为4.86,与家蚕β型微管蛋白1,2,3和4的相似性分别为97%,97%,87%和93%。系统进化分析表明,克隆到的菜粉蝶微管蛋白基因属β型微管蛋白。RT-PCR分析表明,蝶蛹金小蜂寄生能抑制肌动蛋白、肌动蛋白解聚因子及微管蛋白基因在菜粉蝶蛹血细胞中的转录水平。由此推断蝶蛹金小蜂调控寄主血细胞骨架相关蛋白基因的转录是该蜂抑制寄主血细胞免疫反应的分子机制之一。
To reveal the molecular mechanism by which Pteromalus puparum inhibits the hematopoietic response of Pieris rapae, the actin, actin depolymerizing factor, and tubulin cDNA were cloned. The effects of Pteromalus puparum The effect of parasitism on its transcriptional level. The results showed that the cDNA ORF of Actinine was 1131bp, encoding 377aa. The predicted molecular weight was 41.78kDa and the pI was 5.29. The similarity to other insect actin was very high at above 90%. Amino acid composition and phylogenetic analysis showed that the cloned Apante actin gene belongs to cytoplasmic actin. The cDNA of actin depolymerization factor was 1243bp in length and ORF was 447bp, encoding 149aa. The predicted molecular weight was 16.97kDa and the pI was 7.11. The results showed that the actin depolymerization factor cDNA was found to bind to Bombyx mori, Tribolium castaneum, Apis mellifera, The similarities of the scale-deprived Maconellicoccus hirsutus actin depolymerizing factor were 97%, 87%, 89% and 72%, respectively. The cDNA of canola tubulin cDNA was 1757bp in length and 1344bp in ORF, encoding 448aa. The predicted molecular weight was 50.38kDa and the pI was 4.86. The similarity to the β-tubulin 1, 2, 3 and 4 was 97% 97%, 87% and 93%. Phylogenetic analysis showed that the cloned canola tubulin gene belongs to β-tubulin. RT-PCR analysis showed that parasitism of Coprinus copeola inhibits the transcriptional level of actin, actin, and tubulin genes in the blood cells of the young butterfly pupal pupae. Therefore, it was concluded that the transcription of the cytoskeleton-related protein gene of the C. pipiens pallens was one of the molecular mechanisms that suppress the host immune responses of the host cells.