甜菜多粘菌基因组片段的克隆及PCR检测  

THE CLONING AND PCR DETECTION OF GENOMIC DNAFRAGMENT OF POLYMYXA BETAE

作　　者：王琦[1] 韩成贵[1] 李大伟[1] 于嘉林[1] 王慧敏[1] 蔡祝南[1] 刘仪[1] 

机构地区：[1]中国农业大学植物病理系,北京100094

出　　处：《菌物系统》1999年第4期436-439,共4页Mycosystema

摘　　要：根据资料报导设计引物，扩增Ｐｏｌｙｍｙｘａ　ｂｅｔａｅ的基因组片段，将其克隆在ｐＧＥＭ－３Ｚｆ（＋）质粒载体上，并通过双酶切、ＰＣＲ扩增和部分序列测定，证明克隆片段为Ｐ．ｂｅｔａｅ基因组片段。用扩增Ｐ．ｂｅｔａｅ基因组片段的引物对由Ｐ．ｇｒａｍｉｎｉｓ侵染小麦和Ｏ．ｂｒａｓｓｉｃａｅ侵染豇豆根系抽提的总ＤＮＡ进行ＰＣＲ扩增，均未获得任何ＤＮＡ产物，进一步证实了上述结果。对移入病土２、３．５天的甜菜苗单株根系进行ＤＮＡ粗提，移入病土３．５天的甜菜苗ＰＣＲ检测其已被Ｐ．ｂｅｔａｅ侵染，对确定Ｐ．ｂｅｔａｅ的早期侵染有重要意义。According to reports, primers were designed to amplify the genomic DNA fragment of P. betae,which was cloned to pGEM--3Zf(+). By restriction endonuclease digestion, PCR amplification and sequenceanalysis, the cloned DNA fragment is identificated as gemomic fragment of P. betae. NO any DNA product wasobtained in detection of P. graminis or O. brassicae from infected roots of wheat or string bean respectively,which further proved that the primers used for DNA fragment amplification of P. betae is specific to P. betaeonly. Posttransplant in infected soil for two or thee and half a days, cruds DNA was extracted from single rootof sugar beet seedlings and used for PCR test which showed that P. betae DNA could be identified specificallyin the sugar beet seedlings with easy infection period.

关 键 词：甜菜多粘菌 PCR检测 基因组 克隆 

分 类 号：Q949.310.3[生物学—植物学] Q78