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Expressional analysis of the silkworm storage protein 1 and identification of its interacting proteins.
Insect Molecular Biology ( IF 2.6 ) Pub Date : 2019-07-30 , DOI: 10.1111/imb.12610
Ruilin Li 1, 2 , Congwu Hu 1, 2 , Tao Geng 3 , Dingding Lv 4 , Kun Gao 1, 2 , Xijie Guo 1, 2 , Chengxiang Hou 1, 2
Affiliation  

Storage proteins are haemolymph‐specific proteins in insects, mainly synthesized in the fat body, released into the haemolymph, and then selectively reabsorbed by the fat body before pupation. These storage proteins play an important role in insect metamorphosis and egg development. Some of these storage proteins are responsive to pathogen infection and can even suppress pathogen multiplication. However, the mechanisms of the physiological, biochemical and immune‐responsive functions of storage proteins remain unclear. In this study, the expression patterns of Bombyx mori storage protein 1 (BmSP1) during the larval stage were analysed. Then, BmSP1 protein fused with enhanced green fluorescent protein (EGFP) was successfully expressed in a B. mori baculovirus vector expression system. Quantitative real‐time PCR showed that the expression level of BmSP1 increased with the advance of instars and reached the highest level in the fifth instar, especially in the fat body. Recombinant BmSP1 expressed in silkworm larvae inhibited haemolymph melanization. Then, proteins that interact with BmSP1 were identified with EGFP used as an antigenic determinant by co‐immunoprecipitation. A 30 kDa low molecular weight lipoprotein PBMHP‐6 precursor (BmLP6) was shown to interact with BmSP1. Yeast two‐hybrid experiments confirmed the interaction between BmSP1 and BmLP6. The results obtained in this study will be helpful for further study of the functions of BmSP1 and BmLP6 in the regulatory network of silkworm development and innate immunity.

中文翻译:

家蚕贮藏蛋白1的表达分析及其相互作用蛋白的鉴定。

贮藏蛋白是昆虫中血淋巴特异的蛋白,主要在脂肪体内合成,释放到血淋巴中,然后在化ation之前由脂肪体选择性地重新吸收。这些存储蛋白在昆虫的变态和卵的发育中起着重要的作用。这些存储蛋白中的一些对病原体感染有反应,甚至可以抑制病原体繁殖。然而,存储蛋白的生理,生化和免疫应答功能的机制仍不清楚。本研究分析了家蚕家蚕贮藏蛋白1BmSP1)在幼虫期的表达模式。然后,与增强型绿色荧光蛋白(EGFP)融合的BmSP1蛋白成功在桑蚕中表达杆状病毒载体表达系统。实时定量PCR显示,BmSP1的表达水平随着龄期的增加而增加,并在第五龄期达到最高,尤其是在脂肪体内。蚕幼虫中表达的重组BmSP1抑制了血淋巴的黑色素化。然后,用EGFP通过共免疫沉淀法鉴定了与BmSP1相互作用的蛋白。30 kDa的低分子量脂蛋白PBMHP-6前体(BmLP6)与BmSP1相互作用。酵母双杂交实验证实了BmSP1和BmLP6之间的相互作用。这项研究获得的结果将有助于进一步研究BmSP1和BmLP6在家蚕发育和先天免疫调节网络中的功能。
更新日期:2019-07-30
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