Objective      To construct a recombinant lentiviral vector carrying RPS14 gene

establish a stable packaging 293T cell lines

and observe RPS14 expression in human MDS cell line MUTZ-8.     Methods      RPS14 was obtained using reverse transcription polymerase chain reaction (RT-PCR). Then the RPS14 gene was cloned into a lentiviral vector pGC-FU to construct a recombinant lentiviral vector carrying RPS14 gene named pGC-FU RPS14

which was confirmed by PCR

restriction enzyme digestion and DNA sequencing. With the help of Lipofectamine 2000

293T cells were cotransfected with pGC-FU RPS14

pHelper 1.0 and pHelper 2.0. The titer of the virus was examined by real-time PCR. pGC-FU RPS14 was used to transfect human MDS cell line MUTZ-8. The transfection efficiency and RPS14 mRNA expression were detected by flow cytometry and RT-PCR

respectively.     Results      The recombinant lentiviral vector pGC-FU RPS14 was confirmed to be correctly constructed by PCR

restriction enzyme digestion and DNA sequencing; the assayed titer of virus was 2.0×10

9

Tu/ml. The transfection efficiency was 68.41%

and stable expression of RPS14 mRNA in MUTZ-8 cells was detected by RT-PCR.     Conclusion      A lentiviral vector carrying RPS14 gene is successfully constructed

which can deliver RPS14 gene into human MDS cell line MUTZ-8 for RPS14 stable expression.