自我失活型逆转录病毒载体在衰老基因转录调控中的应用

Application of Self-inactivating Retrovirus Vector in Studying Gene Transcription Regulation During The Aging Process

以自我失活型逆转录病毒载体pSIR为基础 ,构建了含有增强型绿色荧光蛋白报告基因的载体pSIR EGFP和含有 870bpp16启动子载体pSIR EGFP 870 .将两种病毒载体转染至包装细胞包装成病毒颗粒 .病毒感染 2BS靶细胞 ,病毒 5′LTR在逆转录过程中自我灭活 .通过该系统观察到 p16启动子在 2BS细胞衰老过程中 ,转录活性明显增强 .结果表明 ,自我失活型逆转录病毒载体能很好地用于衰老基因转录调控的研究 .

Two retrovirus vectors: pSIR-EGFP containing enhance green fluorescence protein (EGFP) reporter gene and pSIR-EGFP-870 with 870 bp p16 promoter were constructed on the base of self-inactivating retrovirus vector pSIR. Then retrovirus vectors were transducted into package cells for virus production and 2BS cells were infected with virus. During the course of reverse transcription, the 5′LTR lost its promoter activity.The transcriptional activity of p16 promoter in 2BS cells obviously increased during the aging process. The results demonstrate that self-inactivating retrovirus vector can be used for gene transcription regulation research during the aging process.