人miR-150相关重组慢病毒载体的包装及其红细胞膜蛋白靶分子的探究
Packaging of recombinant lentiviral vector of miR-150 series and screening of its targets as erythrocyte membrane protein
目的：构建人miR-150相关慢病毒载体并包装慢病毒，探寻其红细胞膜蛋白靶分子。方法：构建miR-150慢病毒载体及阴性对照，并分别包装慢病毒颗粒。同时构建红细胞膜蛋白Band3基因的3'UTR重组质粒psiCHECK2-Band3 3'UTR，将其分别与miR-150 慢病毒或对照慢病毒共转染人胚肾HEK293T细胞，利用荧光素酶报告基因检测二者结合情况。结果：包装成功高滴度的miR-150相关慢病毒颗粒，成功组建psiCHECK2-Band3 3'UTR重组质粒，荧光素酶报告基因显示miR-150能靶向结合Band3的3'UTR。结论：本研究证实了Band3为miR-150的靶标。为miR-150在红系发育过程中的功能和机制研究奠定了实验基础并提供了研究线索。
Object: Constructing hsa-miR-150 related vector and packing the lentivirus and exploring its target belonging to erythroblast membrane protein family. Methods: We construct miR-150 and the negative plasmid and pack the responding lentivirus. At the meanwhile, we construct the recombinant plasmid psiCHECK2-Band3 3'UTR. Then the recombinant plasmid combined with miR-150 lentivirus or the control were co-transfected into 293T cells and the reported assay was employed to detect the binding between miR-150 and Band3 3'UTR. Results: We successfully packed miR-150 related lentivirus with high titer and constructed the recombinant plasmid psiCHECK2-Band3 3'UTR. The reporter assay showed miR-150 was able to bind Band3 3'UTR. Conclusions: This research verified that Band3 is a direct target of miR-150, which lays foundations for studying the roles and related mechanisms of miR-150 in erythropoiesis and also provides clues for further studies.
彭元亮、萧小鹃、孙志卫、曹烨、刘静
基础医学分子生物学生物工程学
分子生物学人miR-150红细胞膜蛋白Band3靶分子
Molecular biologyhsa-miR-150Red blood cell membrane proteinBand3target
彭元亮,萧小鹃,孙志卫,曹烨,刘静.人miR-150相关重组慢病毒载体的包装及其红细胞膜蛋白靶分子的探究[EB/OL].(2016-05-27)[2025-08-02].http://www.paper.edu.cn/releasepaper/content/201605-1452.点此复制
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