摘要
目的 :测定人喉鳞状上皮癌 hep- 2细胞的端粒酶活性水平并获得 hep- 2细胞中 h TERT c DNA片段的克隆。方法 :采用 TRAP- ELISA法测定酶活性水平 ,RT- PCR技术扩增获得目的片段。利用酶切法、PCR法和序列分析法对所获克隆进行检测。结果 :hep- 2细胞具有较高的端粒酶活性水平 ,获得含h TERT c DNA片段的克隆。结论 :利用 RT- PCR法能够从具有较高端粒酶活性水平的 hep- 2细胞中扩增获得 h TERT c
Objective: To assay the telomerase activity level in human laryngocarcinoma cell line hep 2,and to amplify the functional fragment of hTERT (human telomerase reverse transcriptase) cDNA in hep 2.Methods:TRAP ELISA (telomere repeat amplification protocol enzyme linked immunosorbent assay) was used to assay the level of telomerase activity,and RT PCR,to amplify the target fragment.Then,restriction analysis of EcoR Ⅰ and BamH Ⅰ,PCR identification,and DNA sequence analysis were carried out to verify the recombinant plasmids.Results:The telomerase activity level was high in hep 2;the clone carrying the target fragment of hTERT cDNA was obtained.Conclusion:hTERT cDNA can be amplified by RT PCR from hep 2 with high telomerase activity level. [
出处
《白求恩医科大学学报》
CSCD
北大核心
2001年第3期331-333,共3页
Journal of Norman Bethune University of Medical Science
基金
国家自然科学基金资助项目 !(3980 0 10 6 )
