Fei Dong-Qiong, Yu Hai-Zhong, Xu Jia-Ping, Zhang Shang-Zhi, Wang Jie, Li Bing, Yang Li-Ang, Hu Pei, Xu Xin, Zhao Kang, Shahzad Toufeeq
School of Life Sciences, Anhui Agricultural University, Hefei, China; Anhui International Joint Research and Development Center of Sericulture Resources Utilization, China.
School of Life Sciences, Anhui Agricultural University, Hefei, China; Anhui International Joint Research and Development Center of Sericulture Resources Utilization, China; National Navel Orange Engineering and Technology Research Center, Gannan Normal University, Ganzhou, China.
Dev Comp Immunol. 2018 Sep;86:130-137. doi: 10.1016/j.dci.2018.05.012. Epub 2018 May 9.
Ferritin is a ubiquitous iron storage protein that plays an important role in host defence against pathogen infections. In the present study, native ferritin was isolated from the hemolymph of Bombyx mori using native-polyacrylamide gel electrophoresis (native-PAGE) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The results revealed that ferritin consisted of two subunits, designated as BmFerHCH and BmFerLCH. Previously integrated previous transcriptome and iTRAQ data showed that the two subunits were down-regulated in resistant silkworm strain BC9 and there was no obvious change in the expression levels of the subunits in susceptible silkworm strain P50 after BmNPV infection. Virus overlay assays revealed that B. mori ferritin as the form of heteropolymer had an interaction with B. mori nucleopolyhedrovirus (BmNPV), but it can't interact with BmNPV after depolymerisation. What's more, reverse transcription quantitative PCR (RT-qPCR) analysis suggested that BmFerHCH and BmFerLCH could be induced by bacteria, virus and iron. This is the first study to extract B. mori ferritin successfully and confirms their roles in the process of BmNPV infection. All these results will lay a foundation for further research the function of B. mori ferritin.
铁蛋白是一种普遍存在的铁储存蛋白，在宿主抵御病原体感染中发挥重要作用。在本研究中，利用非变性聚丙烯酰胺凝胶电泳（native-PAGE）和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）从家蚕血淋巴中分离出天然铁蛋白。结果显示，铁蛋白由两个亚基组成，分别命名为BmFerHCH和BmFerLCH。先前整合的转录组和iTRAQ数据表明，这两个亚基在抗性家蚕品系BC9中表达下调，而在易感家蚕品系P50中，BmNPV感染后亚基表达水平无明显变化。病毒覆盖分析表明，家蚕铁蛋白以杂聚物形式与家蚕核型多角体病毒（BmNPV）相互作用，但解聚后不能与BmNPV相互作用。此外，逆转录定量PCR（RT-qPCR）分析表明，BmFerHCH和BmFerLCH可被细菌、病毒和铁诱导。这是首次成功提取家蚕铁蛋白并证实其在BmNPV感染过程中的作用的研究。所有这些结果将为进一步研究家蚕铁蛋白的功能奠定基础。
