【摘 要】
：
目的: 构建负载人端粒酶逆转录酶(hTERT)基因的复制缺陷型腺病毒.方法: 将克隆有正义hTERT cDNA的腺病毒穿梭质粒pDC315-hTERT与腺病毒包装质粒pBHGloxDeltaE1,3Cre共转染人
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目的: 构建负载人端粒酶逆转录酶(hTERT)基因的复制缺陷型腺病毒.方法: 将克隆有正义hTERT cDNA的腺病毒穿梭质粒pDC315-hTERT与腺病毒包装质粒pBHGloxDeltaE1,3Cre共转染人胚肾293细胞(HEK 293), 包装负载hTERT基因的复制缺陷型腺病毒表达载体(Ad-hTERT). 对Ad-hTERT扩增、纯化并浓缩后,进一步行感染性鉴定、电镜鉴定及双引物PCR鉴定. 结果: 纯化浓缩后的Ad-hTERT滴度可达5×1012 pfu/L. 电镜下可见在HEK 293细胞中形成的病毒颗粒,PCR双引物鉴定Ad-hTERT可扩增出Ad及hTERT特异片段,而对照则不能扩出hTERT片段. 结论: 成功构建了负载hTERT基因的复制缺陷型腺病毒.
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