反转录病毒载体RCASBP介导的EGFP基因在DF-1细胞中的表达

doi:10.16431/j.cnki.1671-7236.2015.10.004

Abstract

Abstract: To construct a lentiviral vector RCASBP carrying the enhanced green fluorescent protein (EGFP) gene which could be expressed stably in DF-1 cell,EGFP gene was amplified by PCR and then inserted into the lentiviral vector RCASBP after digested with the restriction endonuclease ClaⅠto construct recombinant lentiviral vector RCASBP-EGFP.The recombinant vector was transfected into DF-1 cells by Lipofectamine^TM 2000.Avian leukosis virus (ALV) p27 antigen ELISA test was performed after four passages of the transfected cells and the positive results of ELISA suggested the success rescue of the virus.The expression of EGFP was observed in more than 80 percentages of DF-1 cells under fluorescence microscope.The proviral genome PCR showed EGFP gene carried by the recombinant lentiviral vector RCASBP-EGFP had been integrated into the genome of DF-1 cells.The RCASBP lentiviral-mediated expression system provided a basis for study of the structure and function of ALV genes.

Key words: avian leukosis virus; lentiviral vector RCASBP; enhanced green fluorescent protein (EGFP) gene; DF-1 cell

CLC Number:

Q785

QIN Jian-ru, LI Na-ming, XU Chang-jian, LIAO Ming, CAO Wei-sheng. Expression of Enhanced Green Fluorescent Protein (EGFP) Gene Carried by a Lentiviral Vector RCASBP in DF-1 Cell[J]. , 2015, 42(10): 2538-2543.

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Expression of Enhanced Green Fluorescent Protein (EGFP) Gene Carried by a Lentiviral Vector RCASBP in DF-1 Cell

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