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《Acta Academiae Medicinae Militaris Tertiae》 2006-24
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EBV-LMP1 carboxy region RNA synthesis via transcription in vitro and initial selection of targeted DNAzyme

YANG Yu-cheng,WANG Yi-qin,QIAN Di,HE Yun,HUANG Jiang-ju,HONG Su-ling(Department of Otolaryngology,The First Affiliated Hospital,Chongqing University of Medical Sciences,Chongqing 400016,China)  
Objective To synthesize highly pure EBV-LMP1 carboxy region RNA by in vitro transcription and select its targeted active DNAzyme.Methods EBV-LMP1 exon c gene was amplified by nest PCR from the B95-8 cell genomic DNA and cloned into plasmid pGEM-11zf.The cloned DNA sequence was transcribed in vitro by T7-RNA polymerase.Three targeted 10-23 DNAzymes were designed and synthesized.The highly active and specific DNAzymes were evaluated by in vitro cleavage reaction.Results EBV-LMP1 exon c gene recombinant plasmid was successfully constructed and 40.25 μg EBV-LMP1 carboxy region RNA was produced via transcription in vitro from 1.0 μg lined recombinant plasmid.The cleavage ratio of active DNAzyme DZ1 was 89%.Conclusion Highly pure EBV-LMP1 carboxy region RNA was synthesized successfully by in vitro transcription.One active targeted DNAzyme was selected by in vitro cleavage reaction,paving the way for further research.
【Fund】: 重庆市卫生局资助重点项目(2003)~~
【CateGory Index】: Q789
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