Santra Dipak K, Sandhu Devinder, Tai Thomas, Bhattacharyya Madan K
Department of Agronomy, Iowa State University, Ames, IA 50011, USA.
Funct Integr Genomics. 2003 Dec;3(4):153-9. doi: 10.1007/s10142-003-0092-8. Epub 2003 Oct 1.
We report the construction and characterization of the first soybean yeast artificial chromosome (YAC) library using high-molecular weight DNA isolated from leaf nuclei of the cultivar Conrad 94 that carries Phytophthora resistance genes Rps1-k and Rps6. The quality of this library has been evaluated through analysis of 393 randomly selected YAC clones. The library consists of 36864 clones, of which approximately 19956 carry single soybean YACs with an average size of about 285 kb. The library represents approximately five soybean genome equivalents. The probability of finding any soybean sequences from this library is about 0.99. The library was screened for 43 SSR markers representing the whole soybean genome. We were able to identify positive YAC pools for 95% of the SSR markers. Two YAC clones carrying molecular markers linked to the Rps6 gene were identified. The YAC library reported here would be a useful resource for map-based cloning of agronomically important soybean genes and also to complement the effort towards construction of the physical map for the soybean genome.
我们报道了首个大豆酵母人工染色体（YAC）文库的构建及特性分析，该文库使用从携带疫霉抗性基因Rps1-k和Rps6的Conrad 94品种叶片细胞核中分离的高分子量DNA构建。通过对393个随机选择的YAC克隆进行分析，评估了该文库的质量。该文库由36864个克隆组成，其中约19956个携带单个大豆YAC，平均大小约为285 kb。该文库代表了约五个大豆基因组当量。从该文库中找到任何大豆序列的概率约为0.99。用代表整个大豆基因组的43个SSR标记对该文库进行筛选。我们能够鉴定出95%的SSR标记的阳性YAC池。鉴定出两个携带与Rps6基因连锁的分子标记的YAC克隆。本文报道的YAC文库将是基于图谱克隆重要农艺性状大豆基因的有用资源，也有助于补充大豆基因组物理图谱的构建工作。
