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小麦条锈菌大片段基因组DNA的提取方法研究
引用本文:庄华,岳海梅,郑文明,王晓杰,康振生.小麦条锈菌大片段基因组DNA的提取方法研究[J].菌物学报,2011,30(5):817-821.
作者姓名:庄华  岳海梅  郑文明  王晓杰  康振生
作者单位:1. 西北农林科技大学植物保护学院陕西省农业分子生物学重点实验室 杨凌712100
2. 西藏农牧学院植物科学学院 林芝860000
3. 河南农业大学生命科学学院 郑州450002
基金项目:国家重点基础研究发展规划(973计划)(No. 2006CB100203);现代农业产业技术体系建设专项资金;国家“十一五”支撑计划(No. 2006BAD08A05);教育部长江学者和创新团队发展计划(No. 200558);教育部“111”引智项目(No. B07049)
摘    要:由条锈菌Puccinia striiformis引致的小麦条锈病是小麦最重要的病害之一。由于其活体寄生的特点,对小麦条锈菌的遗传学和分子生物学研究十分有限,大片段核DNA的提取研究还未见报道。高分子量基因组DNA是开展大片段基因组文库构建、基因组分析以及基因组重建的重要基础,通过系统建立和优化小麦条锈菌大片段基因组DNA的分离方法,成功获得分子量大于1Mb高质量的病菌基因组DNA。

关 键 词:小麦条锈菌  高分子量基因组DNA  分离方法

Extraction method of large genomic DNA for wheat stripe rust
Authors:ZHUANG Hu  YUE Hai-Mei  ZHENG Wen-Ming  WANG Xiao-Jie and KANG Zhen-Sheng
Institution:College of Plant Protection, Shaanxi Key Laboratory of Molecular Biology for Agriculture of Northwest Agriculture & Forestry University, Yangling 712100, China;Plant Science Department of Tibet Agricultural and Animal Husbandry College, Linzhi 860000, China;College of Life Sciences, Henan Agricultural University, Zhengzhou 450002, China;College of Plant Protection, Shaanxi Key Laboratory of Molecular Biology for Agriculture of Northwest Agriculture & Forestry University, Yangling 712100, China;College of Plant Protection, Shaanxi Key Laboratory of Molecular Biology for Agriculture of Northwest Agriculture & Forestry University, Yangling 712100, China
Abstract:The stripe rust caused by Puccinia striiformis is one of the most important diseases of wheat. Because of its parasitic living characteristics, studies on genetics and molecular biology of the rust is largely limited. Extraction of large fragments of nuclear DNA from P. striiformis has not been reported, but high molecular weight genomic DNA is very important to construct a large fragment genomic library, and perform sequencing and analysis of genome. In this study, we systematically established and optimized a method of separating large genomic DNA of wheat stripe rust, and over 1Mb of genomic DNA in molecular weight was observed by pulse electrophoresis, indicating that high quality of the genomic DNA was obtained successfully, and could be used for constructing large fragment genomic library and further genomic sequencing of wheat stripe rust.
Keywords:stripe rust pathogen  high molecular weight genomic  separation method
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