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目的构建马尔尼菲青霉菌(Penicilium marneffei, PM)的 LIG4基因敲除载体,为敲除 LIG4基和后期构建 PM 高效打靶系统提供基础.方法通过 PCR扩增 LIG4基因两翼基因片段及筛选标记基因 pyrG 基因片段,再用融合 PCR 方法扩增构建 PM LIG4基因敲除载体,并酶切验证.结果用融合 PCR 成功构建了以pryG 为筛选标记基因的 PM LIG4基因敲除载体.结论马尔尼菲青霉菌 LIG4基因敲除载体构建成功,为进一步同源从组敲除 PM 的 LIG4基因构建高效打靶系统提供基础.“,”Objective: Build Penicilium marneffei (P.marneffei) LIG4 knockout vector to knockout LIG4 gene, providing a foundation for building a highly efficient gene targeting system later. Methods: The LIG4 gene two wings of gene fragments and screening marker gene pyrG gene fragment was amplified by PCR and P.marneffei LIG4 gene knockout vector was amplified by fusion-PCR. Finaly, the gene knockout vector was verification by digestion. Results: P.marneffei LIG4 gene knockout vector was constructed successfuly by using fusion-PCR, which was marked by pryG gene. Conclusions: The P.marneffei LIG4 knockout vector was constructed successfuly, and providing a basis for knocking out LIG4 gene and building a highly efficient targeting system of P.marneffei for further.