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龟纹瓢虫视蛋白基因pj-lwpj-uv1的克隆及表达谱分析
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引用本文:葛亚菲,万岩然,胡雪娇,董丽萍,张煜人,王亚超,范凡,魏国树.龟纹瓢虫视蛋白基因pj-lwpj-uv1的克隆及表达谱分析.植物保护学报,2024,51(6):1393-1402
DOI:10.13802/j.cnki.zwbhxb.2024.2024021
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作者单位E-mail
葛亚菲 河北农业大学植物保护学院, 保定 071000  
万岩然 河北农业大学植物保护学院, 保定 071000  
胡雪娇 河北农业大学植物保护学院, 保定 071000  
董丽萍 河北农业大学植物保护学院, 保定 071000  
张煜人 河北农业大学植物保护学院, 保定 071000  
王亚超 河北农业大学植物保护学院, 保定 071000  
范凡 河北农业大学植物保护学院, 保定 071000 auhff@hotmail.com 
魏国树 河北农业大学植物保护学院, 保定 071000 weiguoshu03@aliyun.com 
中文摘要:为深入研究龟纹瓢虫Propylea japonica视蛋白在视觉系统中的作用,从该虫头部转录组中筛选视蛋白基因进行序列测定及蛋白序列特征分析,并运用实时荧光定量PCR技术分析其在龟纹瓢虫不同发育阶段及成虫不同组织中的表达谱。结果显示:克隆得到龟纹瓢虫的长波敏感视蛋白基因和紫外敏感视蛋白基因序列(GenBank登录号分别为PP356594和PP356595),分别命名为pj-lwpj-uv1,开放阅读框分别为1 122 bp和1 089 bp,分别编码373个和362个氨基酸,均具有7个跨膜拓扑结构,是典型的视觉蛋白。龟纹瓢虫的视蛋白PJ-LW和PJ-UV1分别与鞘翅目瓢甲科昆虫的视蛋白聚集在一个分支,具有高度保守性。龟纹瓢虫pj-lwpj-uv1基因主要在成虫期表达,其表达量均在雌成虫体内最高,分别是雄成虫体内表达量的1.18倍和1.20倍;且均在成虫头部高表达。表明视蛋白基因在龟纹瓢虫成虫视觉系统中具有重要作用,头部和复眼是其发挥视觉功能的关键部位。
中文关键词:龟纹瓢虫  视蛋白基因  基因克隆  序列分析  表达分析
 
Molecular cloning and expression profiling of opsin genes pj-lw and pj-uv1 in the ladybird Propylea japonica
Author NameAffiliationE-mail
Ge Yafei College of Plant Protection, Hebei Agricultural University, Baoding 071000, Hebei Province, China  
Wan Yanran College of Plant Protection, Hebei Agricultural University, Baoding 071000, Hebei Province, China  
Hu Xuejiao College of Plant Protection, Hebei Agricultural University, Baoding 071000, Hebei Province, China  
Dong Liping College of Plant Protection, Hebei Agricultural University, Baoding 071000, Hebei Province, China  
Zhang Yuren College of Plant Protection, Hebei Agricultural University, Baoding 071000, Hebei Province, China  
Wang Yachao College of Plant Protection, Hebei Agricultural University, Baoding 071000, Hebei Province, China  
Fan Fan College of Plant Protection, Hebei Agricultural University, Baoding 071000, Hebei Province, China auhff@hotmail.com 
Wei Guoshu College of Plant Protection, Hebei Agricultural University, Baoding 071000, Hebei Province, China weiguoshu03@aliyun.com 
Abstract:To further investigate the role of opsins in the visual system of the ladybird Propylea japonica, opsin genes were identified from the head transcriptome, and protein sequences were analyzed. The expression profiles of the opsin genes in different developmental stages and adult tissues were analyzed using real-time quantitative PCR. The results indicated that the long wavelength-sensitive opsin gene and ultraviolet-sensitive opsin gene (GenBank accession numbers PP356594 and PP356595, respectively) of P. japonica were successfully cloned and named pj-lw and pj-uv1, respectively. The open reading frames of pj-lw and pj-uv1 were 1 122 bp and 1 089 bp, encoding 373 and 362 amino acids, respectively, with seven transmembrane topological domains, indicating they are typical visual proteins. Phyloge-netic analysis showed that PJ-LW and PJ-UV1 of P. japonica clustered in the same branch as opsins from other species in Coccinellidae, showing high conservation. The pj-lw and pj-uv1 genes were mainly expressed in the adult stage of P. japonica, with expression levels highest in adult female (1.18-fold and 1.20-fold higher than in adult males, respectively). Both genes were most highly expressed in the adult head. These findings suggest that the opsin genes play an essential role in the visual system of P. japonica, with the head and compound eyes being the key regions for visual function.
keywords:Propylea japonica  opsin gene  gene cloning  sequence analysis  expression analysis
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