镰刀菌基因组DNA的抽提及特异片段的扩增分析  

The Analysis of Extraction of Genome of Fusarium and Amplification of Specific Band

作　　者：张宝俊[1] 李志岗[1] 王建明[1] 郭明霞[1] 徐玉梅[1] 

机构地区：[1]山西农业大学农学院,山西太谷030801

出　　处：《山西农业大学学报（自然科学版）》2006年第2期121-124,共4页Journal of Shanxi Agricultural University(Natural Science Edition)

基　　金：山西省留学基金(2004048);山西省自然科学基金项目(20060110080)

摘　　要：实验运用改良的SDS碱裂解法对30株镰刀菌基因组DNA进行了抽提，并对rDNA ITS区域进行了特异性扩增。结果表明：被测菌株DNA片段大于20 kb．其OD260，／OD280大都在1．6～2．0之间，可见用改进的方法抽提的基因组DNA比较完整，纯度较高。实验采用对镰刀菌属有特异性的寡聚核苷酸为引物，对镰刀菌基因组rDNA基因进行扩增，该引物可以扩增出18S、28S部分片段及ITS区域，电泳检测其片段大小在1 041～1 135 bp之间，且特异性良好，为进一步进行RFLP及核苷酸序列分析奠定了基础。Thirty genomic DNA of Fusarium were extracted with improved SDS alkaline lysis method and the ITS district of rDNA was especially amplified, The results showed that the fragment was 20-357～26 421 bp in the isolates and the OD260/OD280 was 1. 6～2. 0. And the genomic DNA extracted with improved SDS method was more integrated and purer. Geneticr DNA of Fusarium was amplified with mononucleotide as the primer. 18S and 28S rDNA of Fusarium and ITS region were amplified. It also was showed that the PCR amplification fragment was 1 041～ 1 135 bp after electrophoresis examination and had good speciality. And it could be used in RFEP and nucleotide acid sequence analyses.

关 键 词：镰刀菌 基因组DNA PCR扩增 

分 类 号：Q781[生物学—分子生物学]