AFLP指纹图谱技术在动物物种鉴定中的应用.pdfVIP

第32卷 第2期 南京林业大学学报(自然科学版) VoI．32．No．2 2008年3月 Journal of Nanjing Forestry University(Natural Sciences Edition) Mar．，2008 AFLP指纹图谱技术在动物物种鉴定中的应用 林建新 ，成志恒 ，张曼芳 ，俞文君 (1、南京森林公安高等专科学校侦查系，江苏 南京 2l0046；2．中国科学院上海生物工程研究中心，上海 200233) 摘要：用AFLP(扩增片段长度多态性)方法来构建羚牛、家牛和黑熊的DNA指纹图谱，并结合国内实验室的具体 情况，避免使用同位素；采用3条技术路线，并灵活应用了两种酶，同时切割和分别在连接寡聚核苷酸接头前后 进行切割：在引物设计时应用3 端有差异的选择性引物进行PCR扩增，同时对实验结果进行比较。结果表明： 两种酶分别在连接寡聚核苷酸接头前后进行切割，然后用单种引物扩增前一种酶切割的片断能获得比较好的结 果。因此，AFLP可用于野生动物DNA分子水平上的鉴定。 关键词：扩增片段长度多态性；动物物种鉴定；羚羊；家牛；黑熊 中图分类号：Q953 文献标识码：A 文章编号：1000—2006(2008)02—0142—03 Application of AFLP fingerprinting in animal species identification LIN Jian—xin ，CHENG Zhi—heng ，ZHANG Man—fang ，Yu Wen—jun (1、Department of Investigation Nanjing Forest Police College，Nanjing 210046，China；2．Shanghai Research Centre of Biotechnology Chinese Academy of Science，Shanghai 200233，China) Abstract：Takin，Cattle，Asiatic black bear DNA fingerprinting was constructed with AFLP(Amplified Fragment Length Polymorphism)and with no isotope in our laboratory．The total genomic DNA was cut at the same time and before or after ligation of oligonucleotide adapters with two restricted enzymes in three technique methods．The PCR amplication was achieved by the use of 3 end selective primers that extended into the restriction fragments．The results were discussed at the same time．The genomic DNA was cut before and after ligation of oligonucleotide adapters with two restriction enzymes． Then the better amplified results could be achieved by using of one primer that extended into the first restriction fragments． So AFLP could be used for DNA fingerprint identification in wild animal species． Key words：AFLP；Animal species identification；