拟南芥profilin2启动子5′端缺失对维管束特异表达的影响

用 ＰＣＲ法扩增并克隆了 ｐｒｏｆｉｌｉｎ２全长启动子（１６６７ ｂｐ），经 ５’端不同长度缺失，与 ｇｕｓ（ｕｉｄＡ）基因连接，构建植物表达载体，转化伽蓝菜，证实在转基因植株中全长启动子Ｐｆｎ１．７呈维管束特异表达．５’端缺失分析显示，可将该启动子分为 ３个区段：区段 １，－１６６７—１３８０ ｂｐ，缺失该区段后 ｇｕｓ基因由维管束特异表达变为组成型表达，推测在该区段中存在维管束特异表达元件；区段 ２，－１１５３～－５９７ ｂｐ，强烈抑制 ｇｕｓ基因的表达；区段 ３，－５９７～－１ｂｐ，可认为是ｐｒｏｆｉｌｉｎ２的基本启动子．

Effect of 5'-deletion of Arabi-dopsis profilin2 promoter on its vascular specific expression

Based on the published sequence of profilin2 promoter of Arabidopsis thaliana, a full-length promoter (1667 bp) was amplified by PCR. The 5’ -end deletion fragments with length of 1380, 1153, 969 and 597 bp were then fused with gus (uid\) gene respectively. Constructed plant expression vectors were individually transferred into Kalan-choe laciniata and transgenic plants regenerated. GUS his-tochemical assay confirmed that the full-length promoter Pfnl.7 was vascular-specific. Deletion assays showed that profilin2 promoter could be divided into three parts. Deletion of fragment 1 ( -1667 -1380 bp) resulted in constitutive expression, suggesting that element(s) responsible for vascular-specific expression might exist in this region. Fragment 2 located at -1153 - -597 bp strongly inhibited gus gene expression. Fragment 3 ( -597 - -1 bp) is considered as a basic domain of profilin2.

Circ_0072088通过靶向miR-223-3p/PFN2轴促进乳腺癌细胞的生长

目的探究环状RNA(circular RNA,circRNA)0072088在乳腺癌(breast cancer,BC)细胞中的生物学功能及其作用机制。方法在公共基因芯片数据库Gene Expression Omnibus(GEO)中下载GSE101123数据集,通过GEO2R分析得到差异基因。通过实时定量聚合酶链反应(quantitative real time polymerase chain reaction,qRT-PCR)检测BC细胞中circ_0072088的表达水平。采用细胞计数试剂盒-8(Cell Counting Kit-8,CCK-8)、划痕愈合实验和Transwell小室法分别检测circ_0072088对BC细胞增殖、迁移和侵袭的作用。通过CircInteractome和TargetScan数据库预测circ_0072088与微小RNA223-3p(micro RNA-223-3p,miR-223-3p)、miR-223-3p与肌动蛋白结合蛋白profilin 2(PFN2)之间的靶向关系,并通过双荧光素酶报告基因实验和RNA结合蛋白免疫沉淀(RNA binding protein immunoprecipitation,RIP)实验验证circ_0072088、miR-223-3p与PFN2之间的靶向关系。结果circ_0072088在BC细胞系中的表达均显著上调。过表达circ_0072088促进BC细胞的增殖、侵袭和迁移,而上调miR-223-3p可削弱此作用。miR-223-3p是circ_0072088的下游靶点,可以被circ_0072088吸附;PFN2是miR-223-3p的靶基因,可以被circ_0072088间接正向调控。结论circ_0072088通过调节miR-223-3p/PFN2轴促进BC细胞的增殖和转移。