黄曲条跳甲酯酶基因片段的克隆及序列分析
Cloning and sequence analysis of the esterase gene fragment of the striped flea beetle, Phyllotreta striolata
郑丽祯1，2**,傅建炜1,2,杨广2,陈小龙2,钟小露2,尤民生2***
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作者单位：1.福建省农业科学院植物保护研究所福州350013; 2.福建农林大学应用生态研究所福州350002
中文关键词：黄曲条跳甲，抗药性，酯酶基因，克隆
英文关键词：Phyllotreta striolata, insecticide resistance, esterase gene, cloning
中文摘要： 黄曲条跳甲Phyllotreta striolata (Fabricius)是十字花科蔬菜的主要害虫之一，其抗药性问题日益严重，克隆黄曲条跳甲的酯酶基因可为其抗性治理提供必要的科学依据。本文利用反转录多聚酶链式反应（RT-PCR）的方法得到一条281 bp的核酸片段，再利用3′-RACE技术获得该基因片段的3′ 端（1 468 bp），编码442个氨基酸。核苷酸序列同源性分析表明，与赤拟谷盗est1基因和杂拟谷盗est1基因的同源性最高，为71%（211/297），与果蝇alphaesterase5的同源性为70％（156/222），氨基酸序列与杂拟谷盗、赤拟谷盗、果蝇的酯酶基因的同源性分别为43%（188/434）、43%（188/434）、33%（144/432）。从BLAST结果可初步推测该片段应为编码黄曲条跳甲酯酶基因的部分核苷酸序列，该序列在GenBank中的登录号为EU166919。
英文摘要： The striped flea beetle (SFB), Phyllotreta striolata (Fabricius) is a major pest of cruciferous vegetables. The resistance of this pest to insecticides has been increasing in recent years. Cloning of the SFB esterase gene may provide a solid and scientific basis for developing strategies to manage pesticide resistance in this species. RTPCR was undertaken to clone one nucleotide fragment of 281 bp, and the 3′RACE used to obtain a 3′sequence (1 468 bp of the fragment, GenBank accession number EU166919) encoded by 442 amino acids. Homological analysis revealed that the obtained fragment was most similar to the esterase1 genes of Tribolium castaneum and T. confusum, with a nucleotide identity of 71% (211/297). The cloned fragment was also 70% (156/222) identical with the alphaesterase5 gene of Drosophila melanoqaster. The amino acid sequence had 43% (188/434), 43% (188/434) and 33% (144/432) similarity to the esterase genes of T.confusum, T. castaneum and D. melanoqaster, respectively. It can therefore be inferred that the obtained fragment should be a partial nucleotide sequence of the SFB esterase gene.