论文部分内容阅读
玉米(Zea mays L.)病害的发生和流行一直是影响其生产的限制因素,玉米南方锈病、粗缩病是我国东南玉米区及黄淮海夏玉米区的重要病害。尽管目前已经开展了对两种病害抗性基因的遗传规律、染色体定位等相关研究,但对这些基因的克隆及功能鉴定等研究进展缓慢,构建抗性材料BAC文库并对抗病基因进行克隆是目前较为有效的方法之一。本研究以黄淮海夏玉米区优良抗病玉米自交系齐319为材料,利用CopyControl pCC1为载体,通过高分子量DNA的提取、部分酶切与大片段DNA的选择、连接转化等多个方面的优化,构建了玉米细菌人工染色体文库。该文库包含270 720个克隆,平均插入片段大小约90 kb,空载率为1.3%,约覆盖玉米基因组10.43倍。文库构建为这些重要功能基因的克隆及比较基因组学研究提供了基础资料。
The occurrence and prevalence of maize (Zea mays L.) diseases have always been the limiting factors affecting its production. Southern corn rust and cunea are the major diseases in the maize area of southeast China and Huang-Huai-Hai summer maize. Although studies have been carried out on the genetic rules and chromosomal location of the two disease resistance genes, the research on the cloning and functional identification of these genes has been slow. The construction of the BAC library of resistant materials and the cloning of the resistant genes Currently one of the more effective methods. In this study, the maize inbred line Qi 319, an excellent disease-resistant maize in Huang-Huai-Hai region, was used as the material. Using CopyControl pCC1 as the carrier, through the extraction of high molecular weight DNA, partial digestion and DNA selection, Optimization, construction of the corn bacterial artificial chromosome library. The library contains 270 720 clones with an average size of about 90 kb and an empty rate of 1.3%, covering about 10.43 fold of the maize genome. Library construction provides the basic information for the cloning of these important functional genes and comparative genomics research.