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菌落PCR技术在重组质粒筛选与鉴定中的应用
Selection and identification of recombinant plasm ids using colony PCR method
【摘要】 应用以菌落为模板的聚合酶链反应(PCR)技术,筛选插有酒类酒球菌苹果酸-乳酸酶基因mleA和苹果酸通透酶基因mleP序列的重组阳性克隆,筛选到的阳性克隆分别扩增到约1600bp及900bp的条带,且与两个目的基因片段大小一致。提取阳性克隆的质粒进一步进行双酶切(EcoR -Kpn )及质粒PCR鉴定,证明结果正确。菌落PCR技术的应用使重组质粒的筛选和鉴定得以优化和简化。
【Abstract】 Colony PCR method were used for selection and identification of the recombinant plasmids containing mleA and mleP from Oenococcus oeni.About 1 600 bp and 900 bp bands were showed respectively,they were the same as the PCR products of the two gene mleA and mleP.These positive clones were used for plasmid extracting and identified by double restriction enzyme digestion (EcoRⅠ and KpnⅠ) and plasmid PCR method.
【基金】 2003年西北农林科技大学专项基金资助项目
- 【文献出处】 西北农林科技大学学报(自然科学版) ,Journal of Northwest Sci-Tech University of Agriculture and Forestry , 编辑部邮箱 ,2004年09期
- 【分类号】Q785
- 【被引频次】15
- 【下载频次】1504