CAJ | 학술논문

目的：构建人端粒酶逆转录酶（hTERT）启动子调控的PUMA基因腺病毒载体（Ad-hTERT-PUMA）及带有增强型绿色荧光蛋白（EGFP）基因的腺病毒载体（Ad-hTERT-PUMA-EGFP）。方法采用一系列的分子生物学技术将 p53正向凋亡调控因子PUMA基因插入hTERT的启动子下游，再将EGFP为分子标记插入到PUMA基因下游，最终将构建好的载体装入腺病毒。双酶切实验证明重组腺病毒载体 Ad-hTERT-PUMA和Ad-hTERT-PUMA-EGFP构建是否成功。结果载体均成功转配腺病毒。转染12 h后，部分HEK293细胞出现了绿色荧光，可维持到72 h。结论成功构建重组腺病毒载体Ad-hTERT-PUMA和Ad-hTERT-PUMA-EGFP。
목적：구건인단립매역전록매（hTERT）계동자조공적PUMA기인선병독재체（Ad-hTERT-PUMA）급대유증강형록색형광단백（EGFP）기인적선병독재체（Ad-hTERT-PUMA-EGFP）。방법채용일계렬적분자생물학기술장 p53정향조망조공인자PUMA기인삽입hTERT적계동자하유，재장EGFP위분자표기삽입도PUMA기인하유，최종장구건호적재체장입선병독。쌍매절실험증명중조선병독재체 Ad-hTERT-PUMA화Ad-hTERT-PUMA-EGFP구건시부성공。결과재체균성공전배선병독。전염12 h후，부분HEK293세포출현료록색형광，가유지도72 h。결론성공구건중조선병독재체Ad-hTERT-PUMA화Ad-hTERT-PUMA-EGFP。
Objective To construt the adenovirus vectors of Ad-hTERT-PUMA-EGFP containing human telomerase reverse transcriptase(hTERT) ,PUMA and enhanced green fluorescent protein (EGFP) .Methods By a series of molecular biology tech-niques ,PUMA gene was inserted into downstream of the hTERT promoter .And then EGFP was inserted into downstream of the PUMA in adenovirus vectors .Finally ,recombinant adenovirus vector ,Ad-hTERT-PUMA-EGFP was assembled into the adenovi-rus .Results Correct adenovirus laoding vectors was verified by viral genome PCR technology .After 12 h of transfection ,HEK293 cells showed green fluorescence in 72 h .Conclusion The correct structure of the adenovirus vectors containing PUMA gene and EGFP gene was identified by restriction enzyme analysis .