Abstract: The aim of this study is to construct the lentivirus expression vector of human CARABIN and to screen the Diffuse Large B Cell Lymphomas (DLBCL) cell SU-DHL-6 for stable expression of CARABIN. ORF of Carabin was amplified by PCR, then the lentiviral vector of over-expression Carabin was constructed using the method of Homologous recombination, confirmed by DNA sequencing and named pLENTI-cGFP-CARABIN. The constructed recombinant vector was co-transfected with psPAX2 and pMD2.G into HEK293T cells to produce the lentiviral particles on the ratio of 3:2:1 by PEI transfection reagent. The lentiviral particles were transduced into SU-DHL-6 cells and the infection efficiency was measured by Fluorescence Microscope. The cells were sorted by the Flow Cytometer (FCM) after 2 weeks. The expression of CARABIN and GFP in the stable cell lines was detected by Western Blot. The lentiviral vector pLENTI-cGFP-CARABIN for overexpression in blood cells was constructed and validated by sequencing. The cell line of stably overexpresiong CARABIN in SU-DHL-6 was confirmed by Western Blot. The over-expression lentiviral vector of human Carabin has been successfully constructed and cell lines for stable expression of CARABIN have been successfully established. This will shed a light on the future study of CARABIN′s function in a variety of lymphoma or leukemia models.

Key words:  CARABIN, over-expression, stable cell line, Diffuse Large B Cell Lymphomas (DLBCL)