CAJ | 학술논문

目的　应用Southern 印迹杂交法（Southern blot）证实转移基因整合至宿主细胞基因组DNA。方法　酚氯仿法提取基因转导的PA317/AIM及K562/AIM细胞基因组DNA，以聚合酶链反应（PCR）扩增转移基因片段；随机引物法标记32P-DNA探针，与经BamHI酶切、琼脂糖凝胶电泳、转移至尼龙膜的基因组DNA进行Southern杂交反应，检测转移基因的整合。结果　PCR反应和Southern blot分析证实转移基因存在于受体细胞基因组DNA中。结论　Southern 印迹杂交法证实转移基因稳定整合至逆转录病毒生产细胞PA317/AIM及靶细胞K562/AIM中。
목적　응용Southern 인적잡교법（Southern blot）증실전이기인정합지숙주세포기인조DNA。방법　분록방법제취기인전도적PA317/AIM급K562/AIM세포기인조DNA，이취합매련반응（PCR）확증전이기인편단；수궤인물법표기32P-DNA탐침，여경BamHI매절、경지당응효전영、전이지니룡막적기인조DNA진행Southern잡교반응，검측전이기인적정합。결과　PCR반응화Southern blot분석증실전이기인존재우수체세포기인조DNA중。결론　Southern 인적잡교법증실전이기인은정정합지역전록병독생산세포PA317/AIM급파세포K562/AIM중。
Objective　To confirm the integration of transduced genes in the genomic DNA of host cells by Southern blot analysis. Methods　The genomic DNA of transduced PA317/AIM and K562/AIM cells were extracted by phenol-chloroform method. The sequences of foreign genes were detected by polymerase chain reaction (PCR). The BamHI-digested DNA resolved by electrophoresis gel was transferred to nylon member and hybridized with the random primer labeled 32　P-DNA probe. The integration of transduced genes is thus determined by so-called Southern blot. Results　The transduced genes were integrated into the genomic DNA of target cells tested by PCR and Southern Blot.Conclusion　The integration of transduced genes in retroviral producing cells PA317/AIM and target cells K562/AIM can be confirmed by Southern blot analysis.