不同诱导条件下家蝇(Musca domestica)三龄幼虫免疫相关基因的表达研究

Expression Analysis of Immune Genes at Different Induction Conditions of Third Instar Larvae of Musca domestica

旨在研究在不同诱导条件下家蝇三龄幼虫先天性免疫基因的表达情况。采用冷刺激、热刺激及革兰氏阴性菌、革兰氏阳性菌和真菌注射诱导12 h后提取家蝇三龄幼虫总RNA。根据GenBank公布的家蝇磷酸甘油醛脱氢酶(GAPDH)基因、攻击素(Attacin)、天蚕素(Cecropin)、防御素(Defensin)、双翅肽(Diptericin)、溶菌酶(Lysozyme)、热休克蛋白(Heat shock protein,HSP)及家蝇抗真菌肽(MAF-1)基因序列进行RT-PCR反应,以GAPDH基因为内参照,分析在不同诱导条件下家蝇三龄幼虫先天性免疫基因的表达情况。结果显示,不同条件诱导下,家蝇免疫相关基因表达显现较大差异,微生物诱导比物理刺激诱导后家蝇免疫相关基因表达水平高;真菌、阳性菌诱导后家蝇免疫相关基因表达量最高,阴性菌次之;冷刺激诱导最低。微生物及物理刺激均能激活家蝇的免疫系统,在不同条件刺激下,家蝇幼虫机体免疫应激反应不同。

We studied on the innate immune gene expression at different induction conditions of third instar larvae of Musca domestica. We chose the cold, heat stimulation and gram negative bacteria, gram positive bacteria and fungi induced the third instar larvae, 12 hours after the total RNA was extracted from third instar larvae.We designed the primers according to the published GenBank glyceraldehyde phosphate dehydrogenase（GAPDH）, Attacin, Cecropin, Defensin, Diptericin, Lysozyme, heat shock protein（HSP）and Musca antifungal peptide（MAF-1） gene sequences of housefly. We were using the GAPDH as reference gene. Analysed the gene expression in different inducing conditions of third instar larvae of housefly through RT-PCR reaction. Results showed that in different inducing conditions, the genes related innate immunity expression showed large differences. Microbial stimulation showed the higher levels of gene expression than physical stimulation. The gene expression induction of fungi and positive bacteria was the highest, the cold stimulation induced the lowest. The innate immune system of the housefly larvae could activate by both microbial and physical stimulation. In different conditions of stimulation, the body’s immune response to different stress.