抑制牛整合素β5基因表达的shRNA的筛选  

Screening of shRNA for Inhibiting Expression of Integrin β5 Gene in Bos Taurus

作　　者：王浩然[1,2] 王洪梅[2] 朱其军 武建明[2] 刘晓[2] 宋玲玲[2] 何洪彬[2] 王立群[1] 

机构地区：[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

出　　处：《家畜生态学报》2012年第2期20-24,共5页Journal of Domestic Animal Ecology

基　　金：国家自然科学基金(31072160);泰山学者海外特聘专家(何洪彬);国家转基因重大专项(2009ZX08007-006B;2011ZX08007-002);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(201004027;201102034)

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-shR-NA145、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。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.

关 键 词：整合素β5 真核表达 SHRNA WESTERN BLOT 

分 类 号：S811.6[农业科学—畜牧学]