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水稻螟虫(二化螟)的兰尼碱受体基因:分子克隆、可变剪接及表达谱分析

Ryanodine receptor genes of the rice stem borer, Chilo suppressalis: Molecular cloning, alternative splicing and expression profiling.

作者信息

Peng Y C, Sheng C W, Casida John E, Zhao C Q, Han Z J

机构信息

Education Ministry Key Laboratory of Integrated Management of Crop Diseases and Pests, College of Plant Protection, Nanjing Agricultural University, Nanjing 210095, PR China.

Environmental Chemistry and Toxicology Laboratory, Department of Environmental Science, Policy, and Management, University of California, Berkeley, CA 94720-3112, USA.

出版信息

Pestic Biochem Physiol. 2017 Jan;135:69-77. doi: 10.1016/j.pestbp.2016.06.002. Epub 2016 Jun 8.

Abstract

The ryanodine receptor (RyR) of the calcium release channel is the main target of anthranilic and phthalic diamide insecticides which have high selective insecticidal activity relative to mammalian toxicity. In this study, the full-length cDNA of Chilo suppressalis RyR (CsRyR) was isolated and characterized. The CsRyR mRNA has an open reading frame (ORF) of 15,387bp nucleotides, which encodes 5128 amino acids with GenBank ID: KR088972. Comparison of protein sequences showed that CsRyR shared high identities with other insects of 77-96% and lower identity to mammals and nematodes with only 42-45%. One alternative splicing site (KENLG) unique to Lepidoptera was found and two exclusive exons of CsRyR (I /II) were revealed. Spatial and temporal expression of CsRyR mRNA was at the highest relative level in 3rd instar larvae and head (including brain and muscle), and at the lowest expression level in egg and fat body. The expression levels of whole body CsRyR mRNA were increased remarkably after injection of 4th instar larvae with chlorantraniliprole at 0.004 to 0.4μg/g. This structural and functional information on CsRyR provides the basis for further understanding the selective action of chlorantraniliprole and possibly other diamide insecticides.

摘要

钙释放通道的兰尼碱受体(RyR)是邻氨基苯甲酸酯类和邻苯二甲酰胺类杀虫剂的主要作用靶点,这类杀虫剂相对于哺乳动物毒性具有高选择性杀虫活性。在本研究中,分离并鉴定了二化螟RyR(CsRyR)的全长cDNA。CsRyR mRNA具有一个15387bp核苷酸的开放阅读框(ORF),其编码5128个氨基酸,GenBank登录号为:KR088972。蛋白质序列比较表明,CsRyR与其他昆虫具有77%-96%的高度同源性,而与哺乳动物和线虫的同源性较低,仅为42%-45%。发现了一个鳞翅目特有的可变剪接位点(KENLG),并揭示了CsRyR的两个独特外显子(I/II)。CsRyR mRNA的时空表达在3龄幼虫以及头部(包括脑和肌肉)中相对水平最高,而在卵和脂肪体中表达水平最低。用0.004至0.4μg/g的氯虫苯甲酰胺注射4龄幼虫后,全身CsRyR mRNA的表达水平显著增加。关于CsRyR的这种结构和功能信息为进一步理解氯虫苯甲酰胺以及可能其他双酰胺类杀虫剂的选择性作用提供了基础。

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