Malonek S, Meinhardt F
Institut für Mikrobiologie, Westfälische Wilhelms-Universität Münster, Germany.
Curr Genet. 2001 Sep;40(2):152-5. doi: 10.1007/s002940100236.
Conidia of the phytopathogenic fungus Calonectria morganii were transformed to hygromycin B resistance using the hph gene of Escherichia coli as the selective trait, governed by a heterologous fungal promoter and the Aspergillus nidulans trpC terminator. Agrobacterium tumefaciens-mediated transformation yielded stable hygromycin B-resistant clones (average number (106 per 10(7)) [corrected] conidia). Putative transformants appeared to be mitotically and meiotically stable. The presence of the hph gene was checked by PCR. In four randomly chosen transformants, single-copy integrations of the marker gene at different chromosomal sites were proven by Southern analysis.
利用大肠杆菌的潮霉素B抗性基因(hph基因)作为选择标记,在异源真菌启动子和构巢曲霉trpC终止子的调控下,对植物病原真菌摩根拟丽赤壳菌的分生孢子进行转化,使其获得潮霉素B抗性。根癌农杆菌介导的转化产生了稳定的潮霉素B抗性克隆(平均每10⁷个分生孢子有10⁶个抗性克隆)。推定的转化体在有丝分裂和减数分裂中似乎都是稳定的。通过PCR检测hph基因的存在。通过Southern分析证实,在四个随机选择的转化体中,标记基因在不同染色体位点上进行了单拷贝整合。