机构：[a]Bayi Children's Hospital, Beijing 100700, China首都医科大学附属北京儿童医院[b]Department of Pediatrics, First Hospital of Sun Yat-sen University, Guangzhou 510080 Guangdong Province, China儿科儿科科室中山大学附属第一医院[c]Laboratory Medicine Center, Nangfang Hospital of Southern Medical University, Guangzhou 510515 Guangdong Province, China[d]Laboratory of Symptomatology, College of Traditional Chinese Medicine, Southern Medical University, Guangzhou 510515 Guangdong Province, China
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摘要：
Aim: RNAi effect is closely correlated with gene transfer. In this study, we constructed pENTR™/U6-TF-shRNA vector on human tissue factor, so as to lay a foundation for treating blood clotting diseases. Methods: The experiment was performed at Laboratory of Symptomatology, College of Traditional Chinese Medicine, Southern Medical University from November 2006 to May 2007. Human tissue factor (NM-001993) was designed by on-line designer software on Corp. Invitrogen, and shRNA sequence was synthesized, then double strands oligonucleotide (ds oligo) was synthesized and cloned into the pENTR™/U6 plasmid. Connection product was inverted competent cells and multiplied. Plasmid was extracted and sequenced. Results: After renaturation, clear ds oligo and ss oligo were observed under ultraviolet after 4% agarose gel electrophoresis in the synthesized double strands oligonucleotide. Sequencing results indicated that pENTR™/U6-RelB-shRNA plasmid was positive clone. Conclusion: Human tissue factor gene pENTR™/U6-TF-shRNA vector is constructed successfully. It develops stable transfection vector for the application of tissue factor gene RNAi vector in treatment blood clotting diseases.
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中文
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推荐引用方式(GB/T 7714)：
Tang W,Zhuang S.-Q,Yang H.-L,等.Construction and identification of RNAi vector pENTR™/U6-TF-shRNA vector on human tissue factor[J].2008,12(2):
