Characterization and functional analysis of serpin-28 gene from silkworm, Bombyx mori
Characterization and functional analysis of serpin-28 gene from silkworm, Bombyx mori
复制标题
家蚕丝氨酸蛋白酶抑制剂28基因的鉴定及功能分析
DOI：
10.1016/j.jip.2018.10.013
复制
发表时间：
2018-11-01
影响因子：
3.4
通讯作者：
Wei, Guoqing
中科院分区：
文献类型：
--
作者：
Gao, Qiuping;Yang, Liangli;Wei, Guoqing
丝氨酸蛋白酶抑制剂（Serpins）是一个广泛分布的蛋白质超家族，具有SERPIN结构域并参与多种免疫应答。在这项研究中，serpin-28基因被确定在B。研究了桑色素及其在免疫调节中的作用。该基因的开放阅读框为1065 bp，编码354个氨基酸残基的多肽，含有一个SERPIN结构域，预测分子量为40.3 kDa。在大肠杆菌中表达重组Bmserpin-28蛋白，制备兔抗Bmserpin-28多克隆抗体。实时荧光定量PCR分析表明，Bmserpin-28在所有检查的组织中表达，在脂肪体和丝腺中的表达量最大。不同发育阶段的表达谱表明，蛹期表达量最高，卵期表达量最低。用大肠杆菌、白僵菌、藤黄微球菌和B. mori核型多角体病毒），研究了Bmserpin-28在脂肪体和血细胞样品中的表达模式。病原体攻击后，在两种测试组织中记录到Bmserpin-28表达水平的显著上调。此外，Bmserpin-28的RNA干扰导致抗菌肽基因的显著上调。综上所述，我们的结果表明Bmserpin-28可能参与了B的天然免疫。森。
Serine protease inhibitors (Serpins) are a broadly distributed superfamily of proteins with a SERPIN domain and participate in several immune responses. In this study, a serpin-28 gene was identified in B. mori and its role in immune regulation was investigated. This gene has an open reading frame of 1065 bp that encodes a 354-amino acid residue polypeptide containing one SERPIN domain with a predicted molecular weight of 40.3 kDa. Recombinant Bmserpin-28 protein was expressed in Escherichia colt and used to raise rabbit anti-Bmserpin-28 polyclonal antibodies. Quantitative real-time PCR analysis revealed that Bmserpin-28 was expressed in all examined tissues, with maximum expression in the fat body and silk gland. Expression pattern of different developmental stages showed that the highest expression level was in the pupae, while the lowest expression level was recorded at the egg stage. After challenge with four different microorganisms (Escherichia colt, Beauveria bassiana, Micrococcus luteus and B. mori nuclear polyhedrosis virus), the expression pattern of Bmserpin-28 was investigated in fat body and haemocyte samples. A substantial upregulation of Bmserpin-28 expression level was recorded following pathogen challenge in both the tested tissues. Furthermore, RNA interference of Bmserpin-28 resulted in significant upregulation of antimicrobial peptide genes. In summary, our results indicated that Bmserpin-28 may be involved in the innate immunity of B. mori.