Arginine kinase is highly expressed in a resistant strain of silkworm (Bombyx mori, Lepidoptera): Implication of its role in resistance to Bombyx mori nucleopolyhedrovirus
Arginine kinase is highly expressed in a resistant strain of silkworm (Bombyx mori, Lepidoptera): Implication of its role in resistance to Bombyx mori nucleopolyhedrovirus
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DOI：
10.1016/j.cbpb.2010.12.001
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发表时间：
2011-03-01
影响因子：
2.2
通讯作者：
Chen, Keping
中科院分区：
文献类型：
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作者：
Kang, Lequn;Shi, Haifeng;Chen, Keping
利用二维凝胶电泳（2-DE）技术，鉴定了家蚕精氨酸激酶（BmAK）编码基因在家蚕抗核多角体病毒（BmNPV）株NB、NPV易感株306和遗传背景与306相似但耐NPV的近等基因系BC8中差异表达。在本研究中，我们利用RT-PCR和实时定量PCR技术对BmAK的表达谱进行了表征。BmAK在不同发育阶段和不同组织中的表达水平是波动的。值得注意的是，菌株NB和BC8在接种NPV后24小时BmAK的表达量增加了10倍以上，而菌株306的BmAK的基础表达量高于菌株NB和BC8，但这种增加在菌株306中被破坏。针对BmAK的多克隆抗体的Western blotting分析证实了这一观察结果，免疫荧光分析首次表明BmAK主要位于细胞质或细胞质中的某些结构上。这些结果表明，精氨酸激酶参与了家蚕幼虫抗NPV感染的抗病毒过程。(c) 2010爱思唯尔公司版权所有。
A gene encoding Bombyx mori arginine kinase (BmAK) has been indentified differentially expressed in the midguts of Bombyx mori strain NB which is resistant to nucleopolyhedrovirus (BmNPV), strain 306 which is susceptible to NPV and a near isogenic line BC8 with similar genetic background to 306 but resistant to NPV by two-dimensional gel electrophoresis (2-DE). In this study, we characterized the expression profiles of BmAK using RT-PCR and real-time quantitative PCR. The expression level of BmAK fluctuated in various developing stage and various tissue. Remarkably, the expression level of BmAK increased more than 10-fold 24 hours post inoculation (h p.i.) of NPV in strain NB and BC8, while such increment was abraded in strain 306 although the basal expression level of BmAK in strain 306 was higher than that of strain NB and BC8. Western blotting analysis using polyclonal antibody against BmAK verified such observation, and immunofluoresence analysis indicated for the first time that BmAK was mainly located to the cytoplasm or some structures in cytoplasm. These findings suggest that arginine kinase is involved in the antiviral process of Bombyx mori larvae against NPV infection. (c) 2010 Elsevier Inc. All rights reserved.