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Tumor-Specific Expression Detected with the Use of an Expression Vector Driven by Human Telomerase Reverse Transcriptase Gene Promoter
Yin JJ; Fu PF; Wang XB; Jin G(靳刚); Xiu RJ; Xiu, RJ (reprint author), Peking Union Med Coll, Chinese Acad Med Sci, Inst Microcirculat, Beijing 100005, Peoples R China.
Source PublicationJournal of Laboratory and Clinical Medicine
2004
Volume144Issue:6Pages:302-306
ISSN0022-2143
AbstractTo construct the tumor-specific expression vector driven by human telomerase reserve transcriptase gene promoter, we amplified a fragment of the enhanced green fluorescent protein (EGFP) gene from the pEGFP-N 1 plasmid and cloned it into the multiple cloning site of the pLNCX vector, then named the recombinant as pLNCX-EGFP. The fragment of human telomerase reverse transcriptase gene promoter (hTERT) was amplified from the human genome with the use of human telomerase reserve transcriptase gene-specific primers and cloned into the pLNCX-EGFP vector, from which the cytomegalovirus promoter had previously been removed through the use of restriction enzymes, in sense orientation relative to the green fluorescent protein coding sequence. Then the expression vector pLNT-EGFP-under the control of the human telomerase reserve transcriptase gene promoter, which contains green fluorescent protein reporter gene-was successfully constructed. To detect the transcriptional activity of the human telomerase reserve transcriptase gene promoter, we conducted transient transfection of this specific expression vector into human lung fibroblast (HLF) cell lines with high telomerase activity and normal human fetal lung fibroblast (W138) cell lines without telomerase activity. The results of transient transfection showed that the pLNT-EGFP vector strongly expressed the green fluorescent protein reporter gene in telomerase-positive cells but not in telomerase-negative cells.
Subject Area力学
DOI10.1016/j.lab.2004.09.010
Indexed BySCI
Language英语
WOS IDWOS:000225891100004
WOS KeywordCANCER-CELLS ; CLONING ; HTERT
WOS Research AreaMedical Laboratory Technology ; General & Internal Medicine ; Research & Experimental Medicine
WOS SubjectMedical Laboratory Technology ; Medicine, General & Internal ; Medicine, Research & Experimental
Citation statistics
Cited Times:1[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://dspace.imech.ac.cn/handle/311007/15334
Collection力学所知识产出(1956-2008)
Corresponding AuthorXiu, RJ (reprint author), Peking Union Med Coll, Chinese Acad Med Sci, Inst Microcirculat, Beijing 100005, Peoples R China.
Recommended Citation
GB/T 7714
Yin JJ,Fu PF,Wang XB,et al. Tumor-Specific Expression Detected with the Use of an Expression Vector Driven by Human Telomerase Reverse Transcriptase Gene Promoter[J]. Journal of Laboratory and Clinical Medicine,2004,144(6):302-306.
APA Yin JJ,Fu PF,Wang XB,靳刚,Xiu RJ,&Xiu, RJ .(2004).Tumor-Specific Expression Detected with the Use of an Expression Vector Driven by Human Telomerase Reverse Transcriptase Gene Promoter.Journal of Laboratory and Clinical Medicine,144(6),302-306.
MLA Yin JJ,et al."Tumor-Specific Expression Detected with the Use of an Expression Vector Driven by Human Telomerase Reverse Transcriptase Gene Promoter".Journal of Laboratory and Clinical Medicine 144.6(2004):302-306.
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