利用两种不同引物扩增金龟子COⅠ序列片段的研究与对比
Research and comparison of the scarab mitochondrial cytochrome C oxidase Ⅰ (COⅠ) gene fragment sequencecloned by two different primers
田雷雷1,2**席景会2,李克斌1***孙昊雨1,2尹姣1,曹雅忠1
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作者单位：1. 中国农业科学院植物保护研究所植物病虫害国家重点实验室北京100193；2. 吉林大学植物科学学院长春130062
中文关键词：金龟子， COⅠ基因， 特异引物， 通用引物
英文关键词：carab, COⅠ gene, specific primer, universal primer
中文摘要： 利用两类不同的引物，即通用引物（L1490，H2198）与特异引物（Pat，Jerry）分别对4种常见金龟子线粒体细胞色素C氧化酶I（COⅠ）基因片段序列进行扩增和测序，获得长度为689 bp与775 bp的序列。对测序结果进行遗传距离分析，并构建了4种金龟子系统进化树。结果表明，特异引物扩增序列的遗传距离在种内稳定性与种间的差异性都明显优于通用引物扩增序列，利用特异引物扩增序列所构建的系统进化树最符合实际情况，因此利用特异引物扩增序列更能够准确的对金龟子进行分类。
英文摘要： Universal primer (L1490, H2198) and specific primer (Pat, Jerry) were used for cloning the mitochondrial cytochrome C oxidase I (COⅠ) gene fragments of four common scarabs. The homologous sequences were compared and partial nucleotide sequences of COⅠ (689 bp, 775 bp) obtained．The genetic distances of these sequences were analysed and the sequences used to produce three kinds of phylogenetic trees. The results show that the stability and differentials of interspecific distance of the sequences amplified with specific primers are clearly higher than those of sequences amplified with a universal primer. The phylogenetic trees constructed from sequences amplified with the specific primer are more consistent with the current morphological taxonomy of these scarab species. We conclude that sequences amplified with specific primer are more accurate for scarab classification.