摘要
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.
AIM To construct an eukaryotic expression vector for sense and antisense human telomerase RNA components (hTR)gene. METHODS The hTR cDNA fragment contained in the pGRN83 vector from restriction endonuclease digestion was inserted into eukaryotic expression vector pcl-neo in cis-direction or trans-direction using DNA recombinant technology, RESULTS The constructed hTR sense and antisense eukaryotic expression vector(pcl-hTR and pcl-ahTR)was proved to be the same as designed by restriction endonuclease analysis and sequencing. CONCLUSION hTR sense and antisense eukaryotic expression vectors were constructed successfully.This study establishes the basic work for antisense hTR gene therapy for carcinoma and for clarifying the role of hTR telomerase activity regulation.
出处
《世界华人消化杂志》
CAS
2000年第5期491-493,共3页
World Chinese Journal of Digestology
基金
国家自然科学基金
No.39770302~~
关键词
人端粒酶RNA组分
端粒酶
反义基因
胃癌
human telomerase RNA components
eukaryotic expression vector
antisense gene
