摘要
利用随机扩增多态性DNA(RAPD)方法对包括球毛壳ND35在内的20个真菌分离物进行分析,筛选出球毛壳ND35的RAPD扩增特异性DNA片段,经pEASY-T3载体克隆和测序、Seqman整合后,用Primer 5.0软件设计了球毛壳ND35的特异引物对CgND35-1:GTTGCCAGCCCGAGGGGAAG,CgND35-2:GTTACCAGCCATGCCTTGAAG,经优化PCR条件,成功获得球毛壳ND35的SCAR标记,该标记片段长度为329 bp。用CgND35-1/CgND35-2引物对进行了SCAR标记特异性和灵敏度检测,表明该特异引物可用于球毛壳ND35的快速检测。
This paper focused on achieving random amplified polymorphic DNA of 20 fungus isolates including Chaetomium globosum ND35, and transferring them to sequence characterized amplified region （SCAR） markers. Based on the RAPD results of C. globosum ND35, one specific DNA fragment was achieved and recycled, purified, cloned into pEASY -33 vector, sequenced and integrated by software Seqman, then one pair of primers （ CgND35 - 1 ： GTrGCCAGCCCGAGGGGAAG, CgND35 - 2： GTTACCAGCCATGCCTY- GAAG） were designed by software Primer 5.0. With these two primers, the SCAR marker at the length of 329 bp of C. globosum ND35 was amplified successfully by the optimized PCR conditions. It was testified that this pair of specific primers could be used for rapid determination of C. globosum ND35.
出处
《山东农业科学》
2011年第10期8-11,共4页
Shandong Agricultural Sciences
基金
国家自然科学基金资助项目(30872024
30571498)
公益性行业(农业)科研专项项目(201003004)
山东省科技发展计划项目(2010GNC10911)
