Cloning and characterization of a ribonuclease L inhibitor from the silkworm, Bombyx mori
Cloning and characterization of a ribonuclease L inhibitor from the silkworm, Bombyx mori
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DOI：
10.1080/10425170400028871
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发表时间：
2005-02-01
期刊：
影响因子：
--
通讯作者：
Kusakabe, T
中科院分区：
文献类型：
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作者：
Maeda, T;Lee, JM;Kusakabe, T
在病毒感染等事件期间，核糖核酸酶 L (RNase L) 途径在细胞对双链 RNA (dsRNA) 的反应中发挥着重要作用。核糖核酸酶 L 抑制剂 (RLI) 属于 ABC 转运蛋白家族，被称为 RNase L 途径的调节剂。 RLI的同系物在包括果蝇和蚊子在内的许多生物体中都有报道，但它们在昆虫和节肢动物中的功能迄今为止尚未阐明。在本研究中，我们克隆了蚕RLI同源物的cDNA，称为BmRLI，并测定了其核苷酸序列。 RT-PCR分析显示BmRLI mRNA在睾丸、卵巢和脂肪体中有明显表达。从cDNA中，表观分子量为69 kDa的重组蛋白在大肠杆菌和培养的昆虫细胞中表达。虽然上调 BmRLI 表达对 RNAi 没有明显影响，但下调 BmRLI 表达会略微降低 RNAi 效率。
The ribonuclease L ( RNase L) pathway plays an important role in the response of cells to double-stranded RNA ( dsRNA) during the events such as virus infection. Ribonuclease L inhibitor (RLI) belonging to the ABC transporter family is known as a regulator of the RNase L pathway. The homologs of RLI were reported in many organisms including the fruit fly and mosquito, but their functions in insects and arthropods have not been elucidated to date. In the present study, we cloned a cDNA of a silkworm RLI homolog, termed BmRLI, and its nucleotide sequence was determined. RT-PCR analysis revealed that the expression of BmRLI mRNA was marked in the testis, ovary and fat body. From the cDNA, recombinant protein with an apparent molecular mass of 69 kDa was expressed in Escherichia coli and cultured insect cells. Although no obvious effect of up-regulation of the BmRLI expression on RNAi was observed, its down-regulation slightly reduced RNAi efficiency.