Construction of recombinant retroviral vector with rat Islet-1 gene / 吉林大学学报(医学版)

Objective To isolate the rat insulin gene enhancer binding protein 1(Islet-1)gene and construct plEGFP-C1-Islet-1 recombinant retroviral expression vector.Methods The cDNA encoding the rat Islet-1 gene was isolated by RT-PCR method,the cDNA was first cloned into PGET-1 TA vector to facilitate the sequence and then subcloned into the retroviral vector plEGFP-C1.plEGFP-C1-Islet-1 was transfected into PA317 packaging cells with lipofectamine 2000.Transformants were selected in medium containing G418.Results A 1 050 bp DNA fragment was obtained by RT-PCR;plEGFP-C1-Islet-1 recombinant retroviral expression vector was identified by restrictive enzymes digestion,PA317 cells transfected with recombinant vector expressed enhancer green fluorescent protein(EGFP).Conclusion The gene encoding the rat Islet-1 is obtained and plEGFP-C1-Islet-1 expression vector is constructed successfully.

The effect of allogeneic MHC Ⅰ gene modification on the immunologic character in bone marrow cells / 中国免疫学杂志

Objective:To observe the effect of allogeneic MHC Ⅰ gene modification on the immunologic character in bone marrow cells and to investigate the mechanism of induce immunologic tolerance of MHC Ⅰ.Methods:The retroviral expression vector of BALB/C mice H 2D d gene was constructed and transducted into C57BL/6 mice bone marrow cells And the expression of H 2D d on the infected cells was detected by FACS technique Then observe the MLR between the BALB/C mice spleen T cells and the C57BL/6 mice bone marrow cells modified with H 2D d gene by MTT assay The cytotoxic activity of BALB/C mice NK cells to the C57BL/6 mice bone marrow cells modified with H 2D d gene was detected by LDH release assay Results:Either stimulation or response ability of the C57BL/6 mice bone marrow cells to the T cell from BALB/C mice spleen was significantly decreased after modified with H 2D d gene The cytotoxic activity of BALB/C mice NK cells to the C57BL/6 mice bone marrow cells modified with allogeneic MHC Ⅰ gene was significantly lower than which to the C57BL/6 mice bone marrow cells non modified Conclusion:It's maybe a way to induce immunologic tolerance in bone marrow transplantation by the means of modify the bone marrow cells with the MHC Ⅰ gene of the recipient

THE STUDY OF THE DIFFERENTIATION OF NSCs MODIFIED WITH ISLET-1 GENE INTO CHOLINERGIC NEURONS / 解剖学报

Objective To explore whether Islet-1 gene of the rat could induce NSCs to differentiate into cholinergic neurons. Methods NSCs were transduced with Islet-1 recombinant retroviral expression vector.The protein expression of Islet-1 gene in NSCs was detected by immunofluorescence histochemical method.The ability of NSCs to differentiate into ChAT positive cells was observed in vivo and in vitro. Results The numbers of ChAT positive cells were significantly increased in the group of NSCs modified with Islet-1 compared with the control group in vitro.NSCs modified with Islet-1 could differentiate into ChAT positive cells when they were grafted into the corpus striatum of the adult rat brain.Conclusion Islet-1 gene could induce NSCs to differentiate into cholinergic neurons.