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载体构建是利用农杆菌介导建立真菌遗传转化体系的基础。以双元载体pPZP100为骨架,通过限制性内切酶酶切、连接以及去磷酸化将标记基因NptⅡ及StNPS6基因侧翼序列导入目标载体pPZP100中,构建玉米大斑病菌非核糖体肽合成酶6基因敲除载体,并采用冻融法将pPZP100NptⅡNPS6转入农杆菌菌株AGL-1。
Vector construction is the use of Agrobacterium-mediated establishment of fungal genetic transformation system basis. The binary vector pPZP100 was used as the backbone and the flanking sequences of NptⅡ and StNPS6 genes were inserted into pPZP100 by restriction endonuclease digestion, ligation and dephosphorylation to construct the non-ribosomal peptide synthetase 6 gene of S. turcica The vector was knocked down and pPZP100NptIINPS6 was transformed into Agrobacterium strain AGL-1 by freeze-thawing method.