CAJ | 학술논문

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.
목적구건마이니비청매균(Penicilium marneffei, PM)적 LIG4기인고제재체,위고제 LIG4기화후기구건 PM 고효타파계통제공기출.방법통과 PCR확증 LIG4기인량익기인편단급사선표기기인 pyrG 기인편단,재용융합 PCR 방법확증구건 PM LIG4기인고제재체,병매절험증.결과용융합 PCR 성공구건료이pryG 위사선표기기인적 PM LIG4기인고제재체.결론마이니비청매균 LIG4기인고제재체구건성공,위진일보동원종조고제 PM 적 LIG4기인구건고효타파계통제공기출.
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.