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| 抑制牛整合素β5基因表达的shRNA的筛选 | |
| 引用本文： | 王浩然,王洪梅,朱其军,武建明,刘晓,宋玲玲,何洪彬,王立群.抑制牛整合素β5基因表达的shRNA的筛选[J].家畜生态学报,2012,33(2):20-24. |
| 作者姓名： | 王浩然 王洪梅 朱其军 武建明 刘晓 宋玲玲 何洪彬 王立群 |
| 作者单位： | 1.东北农业大学 生命学院，黑龙江 哈尔滨 150030；2.山东省农业科学院 奶牛研究中心，山东 济南 250100； 3.山东省泰安市畜牧兽医局，山东 泰安 271000 |
| 基金项目： | 国家自然科学基金，泰山学者海外特聘专家(何洪彬)，国家转基因重大专项，国家奶牛产业技术体系建设专项经费(何洪彬)，济南市高校院所自主创新计划 |
| 摘 要： | 根据牛整合素β5（Integrin Bata5，ITGB5）的基因序列，设计并构建了真核表达载体pcDNA3.1-flag-β5和shRNA重组慢病毒载体H1-shRNA145、H1-shRNA726，利用脂质体介导法，将pcDNA3.1-flag-β5分别和H1-shRNA145、H1-shRNA726共转染293T细胞，并以针对LacZ基因的H1-shRNA-LacZ 与pcDNA3.1 flag β5共转染293T细胞作为对照，以持家基因β-actin为蛋白内参，通过Western blot检测ITGB5的表达，结果表明，H1-shRNA145、H1-shRNA726均可抑制ITGB5的表达，并有剂量效应，为进一步研究ITGB5的基因功能奠定基础。 |
| 关 键 词： | 整合素β5 真核表达 shRNA Western blot |
| 收稿时间： | 2012-02-06 |
Screening of shRNA for Inhibiting Expression of Integrin β5 Gene in Bos Taurus |
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| WANG Hao-ran , WANG Hong-mei , ZHU Qi-jun , WU Jian-ming , LIU Xiao , SONG Ling-ling , HE Hong-bin , WANG Li-qun.Screening of shRNA for Inhibiting Expression of Integrin β5 Gene in Bos Taurus[J].Acta Ecologae Animalis Domastici,2012,33(2):20-24. | |
| Authors: | WANG Hao-ran WANG Hong-mei ZHU Qi-jun WU Jian-ming LIU Xiao SONG Ling-ling HE Hong-bin WANG Li-qun |
| Institution: | 1. College of Life Science, Northeast Agriculture University,Harbin,Heilongjiang 150030,China； 2. Dairy Cattle Research Center,Shandong Provincial Academy of Agricultural Sciences, Jinan, Shandong 250100,China; 3.Tai'an Bureau of Animal Husbandry, Shandong Province,Tai'an,Shandong 271000,China |
| Abstract: | Based on the sequence of Bos taurus integrin β5(ITGB5),the eukaryotic expression vector pcDNA3.1-flag-β5 and two shRNAs recombinant lentivirus vector H1-shRNA145 and H1-shRNA726 were designed and synthesized.The eukaryotic expression vector pcDNA3.1-flag-β5 was cotransfection 293T cells respectively with H1-shRNA145 and H1-shRNA726 using lipofectamine,and with the LacZ shRNA recombinant lentivirus vector as control.Using the housekeeping gene β-actin as internal reference protein,detected the expression of ITGB5 by Western blot analysis.The results showed that the shRNA recombinant lentivirus vector H1-shRNA145 and H1-shRNA726 can both inhibit the expression of ITGB5 with a dose-response.The research will lay a foundation for further study of the gene function of ITGB5. |
| Keywords: | ITGB5 eukaryotic expression shRNA Western blot |
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