植物SSR 引物开发策略简述.pdfVIP

分子植物育种，2009 年，第7 卷，第4 期，第845-851 页 Molecular Plant Breeding, 2009, Vol.7, No.4, 845-851 新思路、新技术、新方法 Novel Thinking Technology 植物SSR 引物开发策略简述 陈怀琼 隋春 魏建和* 中国医学科学院和北京协和医学院药用植物研究所, 北京, 100193 * 通讯作者, wjianh@263.net 摘 要 SSR 是一种优秀的分子遗传标记，已经广泛应用到遗传多样性检测、遗传种质鉴定、遗传连锁图谱 构建、基因定位与标记辅助育种等多个领域。但SSR 标记应用的前提是根据物种已知DNA 序列设计特异引 物，才能进行PCR 扩增。对于大多数物种而言，目前获得的基因组DNA 及表达的cDNA 序列还非常有限或 者根本没有。由此人们不断的设计出各种方法以开发出尽可能多的SSR 引物，本文简要地介绍了基于序列 数据设计和发展SSR 引物的方法，这对于全基因组序列已经测序的或者已经拥有丰富DNA 数据的物种是 非常快捷的策略。对于那些遗传背景复杂或知之甚少或基因组数据有限的物种，本文也系统的介绍了传统的 文库法、富集法等开发SSR 引物的策略。进一步的本文结合本实验室的研究经验，对FIASCO 磁珠富集法和 ISSR-抑制PCR 法进行了比较。 关键词 SSR, 引物开发策略, 文库法, 富集法 Summary of Strategies for Developing SSR Primer Chen Huaiqiong Sui Chun Wei Jianhe * Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences Peking Union Medical College, Beijing, 100193 * Corresponding author, wjianh@263.net DOI: 10.3969/mpb.007.000845 Abstract Being an excellent molecular genetic marker, SSR has been widely applied in many fields including de- tection of genetic diversity, germplasm identification, genetic linkage map construction, gene mapping and molecu- lar marker-assisted breeding. However, SSR markers are based on species specific primers to amplify DNA se- quence. For most species, the currently available genomic DNA and expression of the cDNA sequence is also very limited or no. Thus people continue to design a variety of ways to develop the largest possible number of SSR primers. This article briefly introduces methods of designing and developing SSR primers based on sequence data, which are effective for species whose whole genome sequence have been sequenced, or already have data-rich DNA species. For nucleic acid sequences unknown or little known species, we also systematically introduce several important methods such as traditional library method and enrichment method. Furthermore, we make a comparison between the FIASCO method and ISSR-suppression PCR method. Keywo