家蚕cyp306a1和bcl-2基因的鉴定及其耐氟性功能研究
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摘要
本研究以家蚕作为模式种,以耐氟品系和敏感品系为材料,应用荧光差异显示技术筛选家蚕耐氟性相关的基因,并对这些差异基因进行功能分析。探索家蚕的耐氟中毒的机制,为设计相关的药物解决当前氟化物对家蚕业的破坏,以及为研究氟化物对人类及其它生物氟中毒的防御和治疗提供新的知识。
其主要结论如下:
1.通过荧光差异显示技术,在家蚕耐氟品系441和敏感品系440中筛选与耐性相关的特异条带,获得了一个P450家族基因Bmcyp306a1。荧光定量分析其在家蚕耐性品系441中肠组织表达量相对较高。推论家蚕Bmcyp306a1可能与耐氟有关。
2.差异显示实验过程中获得了另一个在家蚕耐氟品系高表达基因Bmbcl-2。RT-PCR结果表明,清水对照处理时,Bmbcl-2在家蚕耐氟品系441DZ和感性品系440DZ五龄第48 h,中肠、脂肪体、血液、卵巢和丝腺均有表达,并且在441DZ中表达丰度相对高于440DZ。在添食200 ppm氟化钠后,在耐氟品系441F表达水平明显高于440F品系。推论家蚕Bmbcl-2可能与耐氟有关。
In this research, the fluoride-resistent strain and the sensitive strain were used to identify genes linked to silkworm resistance to fluoride by the fluorescent differential display technique. The mechanism of fluoride was studied to provide the new knowledge on desiging related medicine to solve serious loss in sericicultural industry and protect human and other living thing from fluoride.
The research process and the main conclusions are presented as the following:
1. The fluorescent different display technique-PCR (FDD-PCR) was used to analyze differentially expressed genes in fluoride-resistant silkworm Bombyx mori strain 441 and susceptible strain 440, and a differentially expressed gene, Bmcyp306a1, was obtained. Q-PCR analyzed that Bmcyp306a1 had higher expression level in midgut of strain 441 than 440, which indicated that Bmcyp306a1 may be involved in fluoride resistance by high expression in tissues.
2. Meanwhile, another highly expressed gene, Bmbcl-2, in resistant strain was obtained. RT-PCR results showed that when silkworm was treated with water, Bmbcl-2 expressed in five tissues such as midgut, fat body, lipid ovary and silkgland of strain 441DZ and 440DZ, and its expression level was also higher in 441DZ than that in 440DZ. When silkworm was treated with fluoride, the expression level of Bmbcl-2 was higher in 441F than that in 440F. These results indicated that the high expression level of Bmbcl-2 in resistant stain may be related with fluoride-resistance.
其主要结论如下:
1.通过荧光差异显示技术,在家蚕耐氟品系441和敏感品系440中筛选与耐性相关的特异条带,获得了一个P450家族基因Bmcyp306a1。荧光定量分析其在家蚕耐性品系441中肠组织表达量相对较高。推论家蚕Bmcyp306a1可能与耐氟有关。
2.差异显示实验过程中获得了另一个在家蚕耐氟品系高表达基因Bmbcl-2。RT-PCR结果表明,清水对照处理时,Bmbcl-2在家蚕耐氟品系441DZ和感性品系440DZ五龄第48 h,中肠、脂肪体、血液、卵巢和丝腺均有表达,并且在441DZ中表达丰度相对高于440DZ。在添食200 ppm氟化钠后,在耐氟品系441F表达水平明显高于440F品系。推论家蚕Bmbcl-2可能与耐氟有关。
In this research, the fluoride-resistent strain and the sensitive strain were used to identify genes linked to silkworm resistance to fluoride by the fluorescent differential display technique. The mechanism of fluoride was studied to provide the new knowledge on desiging related medicine to solve serious loss in sericicultural industry and protect human and other living thing from fluoride.
The research process and the main conclusions are presented as the following:
1. The fluorescent different display technique-PCR (FDD-PCR) was used to analyze differentially expressed genes in fluoride-resistant silkworm Bombyx mori strain 441 and susceptible strain 440, and a differentially expressed gene, Bmcyp306a1, was obtained. Q-PCR analyzed that Bmcyp306a1 had higher expression level in midgut of strain 441 than 440, which indicated that Bmcyp306a1 may be involved in fluoride resistance by high expression in tissues.
2. Meanwhile, another highly expressed gene, Bmbcl-2, in resistant strain was obtained. RT-PCR results showed that when silkworm was treated with water, Bmbcl-2 expressed in five tissues such as midgut, fat body, lipid ovary and silkgland of strain 441DZ and 440DZ, and its expression level was also higher in 441DZ than that in 440DZ. When silkworm was treated with fluoride, the expression level of Bmbcl-2 was higher in 441F than that in 440F. These results indicated that the high expression level of Bmbcl-2 in resistant stain may be related with fluoride-resistance.
引文
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