摘要
进行了臂形草内生菌特异DNA片段的克隆及分子鉴定方法的研究。从5个臂形草品种分离的5个内生菌纯培养物中,提取基因组DNA,通过140条随机引物的RAPD分析,引物OPAK10扩增出1条约500bp的共有PCR谱带。此DNA片段命名为BE1。对BE1进行分离、纯化,经点杂交证实BE1为臂形草内生菌特异DNA片段。进一步将BE1片段进行回收、克隆和DNA序列分析。BE1在基因库中进行序列分析,未发现相关同源片段。臂形草内生菌特异DNA片段的获得,为建立一种准确、快速检测臂形草内生菌及特异内生菌方法奠定了分子生物学基础。
Total genomic DNA of5endophyte isolates derived from differential Brachiaria species was extracted.Screened by140ten-base oligonucleotide arbitrary primers,an approximately500bp common PCR fragment (named BE1)was obtained in5endophyte isolates amplified with primer OPAK10.The BE1that was isolated,purified and proved by dot hybridization was found highly associated with endophyte-specific DNA fragment in Brachiaria.BE1was cloned and sequenced.After retrieved in Genebank,it had no similar sequences reported.The results suggest that cloning and characterizing of this endophyte-specific DNA fragment may help develop an accurate method for quick detection of endophytes in Brachiaria.
出处
《热带作物学报》
CSCD
2002年第4期58-61,共4页
Chinese Journal of Tropical Crops
基金
日本政府基金资助
