• 首页关于本刊期刊订阅编委会作者指南过刊浏览
胡超超,张怡,张晨岭,吴云豪,陈婉,李可欣,常青.2018.鸻形目鸟类线粒体基因组测序策略.动物学杂志,53(5):769-780.
鸻形目鸟类线粒体基因组测序策略
Strategy of Amplification and Sequencing of the Mitochondrial Genome of Charadriiformes
投稿时间:2017-12-19  修订日期:2018-08-13
DOI:10.13859/j.cjz.201805012
中文关键词:  鸻形目  线粒体基因组  聚合酶链式反应  通用引物
英文关键词:Charadriiformes  Mitochondrial genome  Polymerase chain reaction, PCR  Universal primers
基金项目:江苏省高校自然科学研究项目(No. 16KJB180003和No. 16KJB180012),江苏开放大学(江苏城市职业学院)“十二五”课题项目(No. 15SEW-Y-018)
作者单位E-mail
胡超超 南京师范大学生命科学学院 南京 210046 huweichen@126.com 
张怡 南京师范大学生命科学学院 南京 210046 954146280@qq.com 
张晨岭 江苏第二师范大学生化系 南京 210013 clzhang@jssnu.edu.cn 
吴云豪 南京师范大学生命科学学院 南京 210046 526567387@qq.com 
陈婉 江苏开放大学环境与生态学院 南京 210036 wanwan0322@163.com 
李可欣 南京师范大学生命科学学院 南京 210046 229774833@qq.com 
常青 南京师范大学生命科学学院 南京 210046 Qingchangnj@163.com 
摘要点击次数: 118
全文下载次数: 98
中文摘要:
      鸻形目(Charadriiformes),全世界约有384个物种,分属于19科94属,种类繁多、分布广泛,是研究迁徙和觅食行为的良好材料。近年来,线粒体基因组的研究快速发展,由于样品难以收集,缺乏系统的测序策略,鸻形目鸟类线粒体基因组的研究相对滞后。引物设计对聚合酶链式反应(PCR)至关重要,一个成功的PCR实验依赖于高质量的特异性引物,本研究拟设计一套用于鸻形目鸟类线粒体基因组扩增的通用引物。对GenBank中现有的鸻形目物种线粒体基因组进行多重比对,发现若干个保守性区域,本研究在该区域设计13对扩增鸻形目鸟类线粒体基因组的通用引物,扩增的目的片段长度均在1.5 kb左右。我们选取4个物种,即灰头麦鸡(Vanellus cinereus)、丘鹬(Scolopax rusticola)、白腰草鹬(Tringa ochropus)和针尾沙锥(Gallinago stenura),进行PCR扩增验证,设计的引物在4个物种中均能顺利扩增、测序,该引物在鸻形目鸟类线粒体基因组扩增中的具有普遍适用性。本研究设计的13对通用引物在扩增鸻形目物种线粒体全基因组中具有较强的应用价值,将为鸻形目的系统发育关系、种群遗传学和生物地理学研究提供珍贵的资源。
英文摘要:
      Charadriiformes (Vertebrate: Aves) is a species-rich order with approximately 384 species belonging to 94 genera of 19 families (Fig. 1), and contains some model species to study migration and foraging behavior. The resources of mitogenome have rapidly accumulated in recent years due to the advanced genomic sequencing, however, Charadriiformes mitogenome has not been well studied (Table 2). Primer design has crucial importance to PCR experiments, and a successful PCR experiment depends on the quality and specificity of the primers. After the multiple sequence alignment, we found many conserved regions. Then, we designed the forward and reverse primers according to the overlapping and conserved regions. In this study, we designed 13 pairs of primers, with the amplified target fragment for each pair about 1.5 kb in length (Fig. 2, Table 1). In order to test the proposed method and prove the performance of these primer pairs, the whole primer settings were applied to amplify and sequence the mitochondrial genomes of four birds: Grey-headed Lapwing Vanellus cinereus, Eurasian Woodcock Scolopax rusticola, Green Sandpiper Tringa ochropus, and Pin-tailed Snipe Gallinago stenura (Fig. 3). The results proved that these primer pairs were efficient. Taken together, the designed primer pairs are an effective toolkit of amplification and sequencing for the Charadriiformes mitogenomes. Our work provides a valuable resource facilitating further study of molecular systematics, population genetics, and phylogeography of Charadriiformes.
附件
查看全文  查看/发表评论  下载PDF阅读器