| 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1 600 bp及900 bp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(EcoRⅠ-KpnⅠ)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。 |
| 关键词: 菌落PCR 重组质粒 苹果酸－乳酸酶基因 苹果酸通透酶基因 |
| DOI： |
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| 基金项目:2003年西北农林科技大学专项基金资助项目 |
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| Selection and identification of recombinant plasmids using colony PCR method |
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| 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 (EcoRⅠand KpnⅠ) and plasmid PCR method. |
| Key words: colony PCR recombined plasmid mleA mleP |
