【作者】 张敏； 张建珍； 郭亚平； 曹天文； 马恩波；

【Author】 ZHANG Min, ZHANG Jian-Zhen, GUO Ya-Ping, CAO Tian-Wen, MA En-Bo College of Life Science and Technology, Shanxi University, Taiyuan 030006, China Institute of Plant Protection, Shanxi Academy of Agriculture Science, Taiyuan 030031, China

【机构】 山西大学生命科学与技术学院； 山西省农业科学院植物保护研究所；

【摘要】 分别用酚-氯仿法和饱和NaCl法对蝶类干标本胸部、腹部和中后足肌肉进行基因组DNA的提取,结果显示:酚-氯仿法所提DNA得率较低,以中后足肌肉为材料提取的DNA在琼脂糖凝胶电泳检测中无可见条带;采用饱和:NaCl法对于制蝶类标本中后足肌肉提取的基因组DNA较完整,得率和纯度也较高,两种方法对胸部和腹部肌肉提取的DNA均出现严重降解。以中后足肌肉提取的基因组DNA为模板,建立RAPD-PCR反应模式,从50条随机引物中筛选出22条可对大紫蛱蝶Sasakia charonda、黑紫蛱蝶Sasakia funebris(Leech)和黑脉蛱蝶Hestina assimilis(Linnaeus)稳定扩增的引物,为开展蝶类分子系统学研究提供了技术手段更多还原

【Abstract】 Genome DNA of butterflies was obtained from the breast muscle, the abdomen muscle and the midleg and hindleg muscle of dry samples using phenol-chloroform protocol and saturated NaCl methods. The results indicated that the yield of DNA extracted was more by using saturated NaCl method than that of phenol-chloroform protocol. The DNA of midleg and hindleg muscle from dry samples of butterflies extracted using phenol-chloroform protocol could not be visualized by ethidium bromide staining of agarose gels, meanwhile the DNA extracted of the midleg and hindleg muscle using saturated NaCl was intact and could fit the need of RAPD-PCR. However, The DNA extracted from the breast and the abdomen muscle using both methods was degraded to a low molecular size. Using the DNA from the midleg and hindleg muscle as the temple, we established RAPD-PCR reaction model. The 50 random primers (10 bp) were analyzed under predetermined optimal reaction conditions, and 22 of them were informative and yielded clear and reproducible bands in Sasakia charonda , Sasakia funebris (Leech) and Hestina assimilis (Linnaeus). These results may provide some references for the further studying the molecular phylogenetic relationships of butterflies更多还原