| Survivin基因及CDK1基因联合靶向shRNA真核表达载体的构建 |
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| 引用本文： | 陈淑萍,符生苗,周红桃,蔡俊宏,李成学,王福利,许茂轩.Survivin基因及CDK1基因联合靶向shRNA真核表达载体的构建[J].海南医学,2014,0(13):1873-1877. |
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| 作者姓名： | 陈淑萍 符生苗 周红桃 蔡俊宏 李成学 王福利 许茂轩 |
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| 作者单位： | 陈淑萍 (海南大学农学院,海南 海口,570228)； 符生苗 (海南省人民医院医学检验中心,海南 海口,570311)； 周红桃 (南方医科大学珠江医院医学检验中心,广东 广州,510280)； 蔡俊宏 (海南省人民医院医学检验中心,海南 海口,570311)； 李成学 (海南省人民医院医学检验中心,海南 海口,570311)； 王福利 (海南省人民医院医学检验中心,海南 海口,570311)； 许茂轩 (海南大学农学院,海南 海口,570228)； |
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| 基金项目： | 国家自然科学基金资助项目(项目编号：编号81060223) |
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| 摘 要： | 目的构建靶向Survivin基因及CDKl基因的短发夹样RNA（Short hairpin RNA，shRNA）真核表达载体。方法根据Genbank报道的Survivin序列及CDKl序列，遵循shRNA设计原则设计并合成各自靶向的Survivin、CDKl基因的shRNA寡核苷酸序列，构建pU6-shRNA-Survivin重组质粒、pu6-shRNA-CDK1重组质粒及pu6-shRNA-Survivin-U6-shRNA-CDK1双基因序列串联重组质粒，并进行限制性内切酶酶切及基因测序鉴定。结果经酶切及测序结果分析，pU6-shRNA-Survivin重组质粒、pu6-shRNA-CDK1重组质粒及pu6-shR-NA-Survivin-U6-shRNA—CDK1双基因系列串联重组质粒均成功构建。结论成功构建Survivin基因及CDKl基因联合靶向shRNA重组质粒，为进一步研究Survivin基因和CDKl基因联合干扰提供了新的方法。
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| 关 键 词： | Survivin基因 CDKI基因 短发夹样RNA pU6-M4 |
Constructing recombinant survivin and CDK1 tandem shRNAs pU6-M4 plasmid |
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| Affiliation: | CHEN Shu-ping, FU Sheng-miao, ZHOU Hong-tao, CAI Jun-hong, LI Cheng-xue, WANG Fu-li, XU Mao-xuan(1. College of Agriculture, Hainan University, Haikou 570228, Hainan, CHINA; 2. Center of Medical Laboratory, People＇s Hospital of Hainan Province, Haikou 570311, Hainan, CHINA; 3. Center of Medical Laboratory, Zhujiang Hospital, Southern Medical University, Guangzhou 510280, Guangdong, CHINA) |
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| Abstract: | Objective To construct recombinant survivin and CDK1 tandem shRNAs pU6-M4 plasmids. Methods Sequences of survivin and CDK1 and the restriction enzyme cutting sites of them were designed and synthesized based on the date from GeneBank. Recombinant plasmids of pU6-M4-Survivin-shRNA and pU6-M4-CDKI-shRNA were constructed and identified. Two determined plasmids were digested by restriction endonucleases then taped by T4 DNA ligase. The ligated products were transformed into competent THSa ceils, and then the recombinant clones were identified by sequencing. Results Restriction enzyme cleave identification and sequencing proved that recombinant survivin and CDK1 tandem shRNAs pU6-M4 plasmids were successfully constructed. Conclusion Recombinant survivin and CDK1 tandem shRNAs pU6-M4 plasmids were correctly constructed. |
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| Keywords: | Survivin Cyclin-dependent kinase 1 Short hairpin RNA pU6-M4 |
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