| 摘要: |
| 【目的】克隆黄曲条跳甲钙离子结合蛋白（reticulocalbin, RCN），并分析其序列特征和表达谱。【方法】结合转录组测序及荧光定量PCR技术，鉴定和分析黄曲条跳甲钙离子结合蛋白基因的序列特征、功能及表达谱。【结果】获得的黄曲条跳甲RCN基因的cDNA序列全长为1 197 bp，开放阅读框为984 bp，共编码327个氨基酸残基。其蛋白分子含有5个亮氨酸拉链结构域（EF-hand），与钙离子结合的模体可能为DX（N/D）X（D/N）XXXXXXE。cDNA序列的系统发育分析表明，黄曲条跳甲的RCN与赤拟谷盗的亲缘关系最近。荧光定量PCR分析表明，RCN在黄曲条跳甲雌雄成虫的不同部位都有表达，具有一定的广谱性，其中在头部和中肠的表达量相对较高；触角中雌虫的相对表达量是雄虫的1.9倍，而在生殖系统中，雄虫的相对表达量是雌虫的2.1倍。【结论】成功鉴定了黄曲条跳甲的一种钙离子结合蛋白基因，初步分析了该基因与钙离子结合的模体序列及在虫体不同部位转录水平的表达情况。 |
| 关键词: 黄曲条跳甲 钙离子结合蛋白 亮氨酸拉链结构域 荧光定量PCR |
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| 基金项目:国家自然科学基金项目（31101500）；仲恺农业工程学院科研基金项目（G3100005） |
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| Cloning,expression analysis of reticulocalbin(RCN) in Phyllotreta striolata(Fabricius) |
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| Abstract: |
| 【Objective】The research identified calcium-binding proteins of Phyllotreta striolata and analyzed its sequence character and gene expression profiles.【Method】EST of calcium binding protein was clustered and identified from data of transcriptome of P.striolata (Fabricius).Gene expression profiles in different tissues were analyzed by real-time PCR.【Result】cDNA of a kind of calcium binding protein,reticulocalbin (RCN) was identified,which contains a 1 197 bp open reading frame (ORF) encoding 327 amino acids.Its putative protein product contains 5 EF-hands and a probably calcium binding motif,DX(N/D)X(D/N)XXXXXXE.Phylogenetic analysis of the complete cDNA sequence showed that the RCN of P.striolata was highly homologous to RCN of Tribolium castaneum.Quantitative real-time PCR assay showed that this RCN mRNA expressed in several tissues,with relatively high expression in head and midgut than that of other organs,including antenna and legs.The expression level of RCN gene in antenna of female adult was 1.9 times as high as that in male.However,the level in production tissue of male was 2.1 times as in female.【Conclution】This study successfully identified a calcium binding protein and analyzed its calcium binding motif and gene expression profiles in different tissues of adult. |
| Key words: Phyllotreta striolata(Fabricius) calcium binding protein EF-hand real-time PCR |
