pMIR-REPORT miRNA  Expression  Reporter Vector熒光素酶報告載體質粒

The  pMIR-REPORT?  miRNA  Expression  Reporter  VectorSystem  consists  of  an  experimental  firefly  luciferase  reportervector  and  an  associated  β-gal  reporter  control  plasmid.  Byinserting  predicted  miRNA  target  sequences  in  the  multiplecloning  site,  pMIR-REPORT  Luciferase  miRNA  ExpressionReporter Vector can be used to conduct accurate, quantitativeevaluations of miRNA function. It can also be used to evaluatesiRNA  target  sites  and  to  analyze  the  influence  of  3' UTRsequences on gene expression. pMIR-REPORT Luciferase con-tains  a  firefly  luciferase  reporter  gene  under  the  control  of  aCMV promoter/termination system (Figure 1). The 3' UTR ofthe luciferase gene contains a multiple cloning site for insertionof  predicted  miRNA  binding  targets  or  other  nucleotidesequences. By cloning a predicted miRNA target sequence intopMIR-REPORT, the luciferase reporter is subjected to regula-tion that mimics the miRNA target.pMIR-REPORT β-gal is a beta-galactosidase reporter plasmidthat is designed for transfection normalization (Figure 2). β-galexpression from this control plasmid can be used to normalizevariability  due  to  differences  in  cell  viability  and  transfectionefficiency. As  an  additional  feature,  pMIR-REPORT  Luciferase  andpMIR-REPORT β-gal include puromycin and neomycin resis-tance genes, respectively, to enable antibiotic selection of trans-fected  mammalian  cells.  Antibiotic  selection  can  be  used  togenerate stable cell lines that express pMIR-REPORT if this isrequired  for  your  research.  For  most  applications,  transientexpression is sufficient, and antibiotic selection is not needed.

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