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目的构建正义和反义端粒酶 RNA 组分((human telomeraseRNA components,hTR)基因真核表达载体.方法用 Mlu Ⅰ和 Sal Ⅰ从 pGRN83质粒上切下约579 bp 的hTR cDNA 片段,分别定向连入 pcl-neo 的 Mlu Ⅰ/Sal Ⅰ和Mlu Ⅰ/Xho Ⅰ酶切位点上,即构建成了 hTR 的正反义表达载体,并经酶切鉴定和测序确认.结果经酶切鉴定和测序证明,所构建的正反义 hTR 真核表达载体与设计完全一致.结论成功构建了人端粒酶 RNA 的正反义真核表达载体,为进一步研究正反义 hTR 基因转染对胃癌细胞端粒酶及生物学行为的影响奠定了基础.
Objective To construct a eukaryotic expression vector for human sense and antisense telomerase RNA components (hTR) gene. Methods Approximately 579 bp hTR cDNA fragment was excised from pGRN83 using Mlu I and Sal I. Into the pcl-neo Mlu I / Sal I and Mlu I / Xho I restriction site, that is constructed hTR positive and antisense expression vectors, and identified by enzyme digestion and sequencing confirmed. The results were identified by enzyme digestion and sequencing It was proved that the constructed eukaryotic expression vector for hTR and antisense hTR was completely identical to the design. Conclusion The positive and antisense eukaryotic expression vectors of human telomerase RNA were successfully constructed to further study the transfection of antisense and hTR genes on gastric cancer cells. The basis of granzymes and biological behavior laid the foundation.