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首页> 外文期刊>International immunopharmacology >Molecular cloning and expression analysis of a hemolin-like molecule from Antheraea pernyi
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Molecular cloning and expression analysis of a hemolin-like molecule from Antheraea pernyi

机译：多年生花An（Antheraea pernyi）苏木素样分子的分子克隆和表达分析

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摘要

Hemolin belongs to the immunoglobulin superfamily and plays an important role in innate immune response of insects. In this study, a hemolin-like cDNA of 1418 bp was obtained from Antheraea pernyi (Ap-hemolin-like). Sequence analysis revealed Ap-hemolin-like was homologous to those hemolins from other insect species. Recombinant Ap-hemolin-like protein was expressed in Escherichia coli cells, and polyclonal antibodies were produced against the recombinant proteins. Real-time PCR and western blot analysis showed that the Ap-hemolin-like was expressed in hemolymph, Malpighian tubules, midgut, epidermis and fat body, with the highest expression level in hemolymph. To investigate its role in the immune response against microorganisms, fifth instar larvae were challenged by injecting nucleopolyhedrovirus (NPV), E. coli, or Beauveria bassiana. The results showed that the expression of Ap-hemolin-like in hemolymph and fat body was obviously-induced by microorganisms. In addition, the recombinant Ap-hemolin-like protein promoted the agglutination of E. coli in the presence of calcium, which was confirmed by agglutination assay. These results suggested that the Ap-hemolin-like protein was involved in innate immune response of A. pernyi against pathogens. (C) 2015 Elsevier B.V. All rights reserved.
机译：血红素属于免疫球蛋白超家族，在昆虫的先天免疫应答中起重要作用。在这项研究中，从per蚕（Antheraea pernyi）（Ap-hemolin-like）获得了1418 bp的helinin-like cDNA。序列分析显示类似Ap-hemolin的蛋白与来自其他昆虫物种的那些血红蛋白是同源的。重组Ap-hemolin样蛋白在大肠杆菌细胞中表达，并产生针对重组蛋白的多克隆抗体。实时荧光定量PCR和Western blot分析表明，Ap-血凝素样蛋白在血淋巴，马氏小管，中肠，表皮和脂肪体中表达，在血淋巴中表达最高。为了研究其在针对微生物的免疫应答中的作用，通过注射核多角体病毒（NPV），大肠杆菌或球孢白僵菌来挑战五龄幼虫。结果表明，微生物明显地诱导了Ap-血凝素样蛋白在血淋巴和脂肪体内的表达。另外，重组Ap-hemolin样蛋白在钙存在下促进了大肠杆菌的凝集，这通过凝集测定法得以证实。这些结果表明Ap-hemolin样蛋白参与了A. pernyi对病原体的先天免疫应答。 （C）2015 Elsevier B.V.保留所有权利。

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