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hTERT核心启动子调控的hNIS重组腺病毒的构建与鉴定 被引量:3

Construction and verification of adenovirus containing the hNIS gene under the control of the hTERT core promoter
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摘要 目的:构建含人端粒酶逆转录酶(hTERT)核心启动子调控的人钠碘转运体(hNIS)基因重组腺病毒,并初步鉴定。方法:将hTERT核心启动子从hTERT/pcDNA3.1(+)质粒中切出,亚克隆至含全长hNIS cDNA序列的质粒载体FL*-hNIS/pcDNA3,应用限制性内切酶将hTERT-hNIS片段切出后连入穿梭质粒载体pAdTrack,应用AdEasy系统构建出重组腺病毒Ad-hTERT-hNIS。同时,构建CMV启动子调控的hNIS重组腺病毒Ad-CMV-hNIS作为阳性对照。应用RT-PCR方法验证hTERT在转染肿瘤细胞中的转录活性。结果:成功构建重组腺病毒Ad-hTERT-hNIS、Ad-CMV-hNIS并经PCR验证正确。RT-PCR证实hNIScDNA能从Ad-hTERT-hNIS转染的细胞中扩增出来。结论:成功构建重组腺病毒Ad-hTERT-hNIS;hTERT核心启动子在转染的肺癌A549细胞中具有转录活性。为下一步的碘治疗实验提供了实验依据。 Objective: To construct a recombinant adenovirus harboring the human sodium iodide symporter (hNIS) under control of the human telomerase reverse transcriptase (hTERT) core promoter. Methods: The hTERT core promoter was cut from plasmid hTERT/pcDNA3.1 ( + ), and then subcloned into plasmid vector FL * -hNIS/pcDNA3 with the full-length hNIS cDNA, and then the hTERT-hNIS sequence was subcloned into the shuttle plasmid pAdTrack. The recombinant adenovirus Ad-hTERT-hNIS was constructed by AdEasy system. In addition, a positive control of adenovirus Ad-CMV-hNIS containing the CMV promoter followed with hNIS gene was created by AdEasy system. The tran scriptional activity of hTERT was examined by RT-PCR in transiently transfected A549 cell lines. Results: Recombinant Ad-hTERT-hNIS was correctly constructed and coni'mned by restriction enzyme analysis and PCR. RT-PCR showed that hNIS cDNA could be amplified from transfected cells. Conclusion: The recombinant adenovirus Ad-hTERT-hNIS is constructed. This study demonstrates that hTERT core promoter has transcriptional activity in transfected A549 cells, and it provides the foundation for further research.
出处 《中国免疫学杂志》 CAS CSCD 北大核心 2008年第6期549-553,共5页 Chinese Journal of Immunology
关键词 钠碘转运体(hNIS) 端粒酶逆转录酶(hTERT) 重组腺病毒 核心启动子 Sodium iodide symporter(hNIS) Telomerase reverse transcriptase(hTERT) Adenovirus Core promoter
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同被引文献14

  • 1贾悦,刘超,覃又文,刘翠萍,唐伟,袁庆欣,狄福松.维甲酸对乳腺癌细胞MDA-MB453钠碘转运体调控(英文)[J].中国现代医学杂志,2004,14(21):11-14. 被引量:2
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