Objective    To construct an eukaryotic expression vector of pre-miR-15a

and to investigate the inhibitory effect of pre-miR-15a to Raji cells.     Methods    The pGCSIL-GFP vector encoding pre-miR-15a nucleotides was transfected into the bacterial competent cells

and then confirmed by PCR and sequencing analysis. The identified vector was transfected into Raji cells with oligofectamine 2000. The cells were divided into 3 groups

blank

negative control and pre-miR-15a group. Semi-quantitative RT-PCR was used to detect the expression of Bcl-2 mRNA

and immunofluorescence indirect for Bcl-2 protein expression. The growth of Raji cells was assayed by trypan blue dye exclusion method.     Results    PCR and sequences analysis indicated that the recombinant clones was identical with target sequences. Many green fluorescent cells were observed under fluorescent microscopy. The levels of Bcl-2 mRNA at every group had no obviously difference. Bcl-2 protein expression was obviously decreased at pre-miR-15a group compared with the other groups. Trypan blue dye exclusion method showed the cell growth was inhibited at 48 h and 72 h post-transfection.     Conclusion    We successfully construct the eukaryotic expression vector of pre-miR-15a

and it can inhibit the growth of Raji cells.