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OALib Journal期刊
ISSN: 2333-9721
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酿酒酵母pdc1基因过表达菌株的构建
DOI: 10.3724/SP.J.1145.2013.00704, PP. 704-708
Keywords: 酿酒酵母,pdc1,基因过表达,荧光定量pcr,乙醇发酵
Abstract:
扩增pdc1全长和kan基因，构建带有kanr和ampr的整合表达载体psh-pdc1，线性化后liac/sscarrierdna/peg法高效转化酿酒酵母ys2-△adh2-△ald6.g418结合pcr筛选获得阳性转化子，然后转入psh65质粒，半乳糖诱导表达cre酶切除kan基因，获得pdc1过表达菌株.荧光定量pcr分析表明过表达菌株pdc1的mrna表达量是出发菌株的2.078倍，遗传性能稳定，生长情况与出发菌株无明显差异.发酵试验显示过表达菌株蔗糖消耗速率较出发菌株缓慢，乙醇最大积累量为72h的12.03%，提高了5.62%.利用同源重组原理和cre-loxp系统，成功构建pdc1基因过表达突变株并提高了乙醇产量.图6表3参17
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