hTERT启动子调控的双顺反子载体的构建
首发时间:2008-01-11
摘要:目的 构建了人端粒酶逆转录酶 (human telomerase reserse transcription, hTERT)基因核心启动子调控的肿瘤细胞特异表达载体。方法 提取人 SGC-7901细胞的基因组 , 利用PCR技术克隆 hTERT基因核心启动子(hTERTp)。并将其插入表达载体pVITRO2中,以hTERTp替换hFerL启动子和hFerH启动子构建hTERTp1-hTERTp2-pVITRO2。用酶切法和测序法, 鉴定重组载体。结果 经酶切和测序鉴定,以hTERT基因核心启动子调控的真核表达载体hTERTp1-hTERTp2-pVITRO2构建成功。结论 hTRET基因核心启动子具有肿瘤特异性, 构建 hTERT基因核心启动子的载体可能是一种新型和有希望肿瘤治疗的途径。
关键词: 端粒酶逆转录酶核心启动子 双顺反子载体pVITRO2 基因治疗
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Construction of the bicistronic vector modulated by hTERT promoter
Abstract:AIM To construct the tumor cell-specific expression vector modulated by human telomerase reserse transcription gene (hTERT) core promoter. METHODS The hTERT gene core promoter fragmentwas amplified by PCR using the totalgenomic DNA from SGC-7901cells as template.The amplified gene fragment was subsequently cloned into pVITRO2 vector,to replace hFerL promoter and hFerH promoter with hTERTp.Identification of hTERTp1-hTERTp2-pVITRO2 by the restriction endonucleases digestion and sequencing.RESULTS The restriction endonucleases digestion and sequencing suggested that the vector hTERTp1-hTERTp2-pVITRO2 were constructed successfully. CONCLUSION The hTERT core gene promoter has the tumor specificity, suggesting that the specific expression vector modulated by hTERT core gene promoter may be a novel and promising approach to the tumor treatment
Keywords: hTERT gene core promoter Bicistronic vector pVITRO2;gene therapy
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No.1801618674812000****
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