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Construction and identification of Kir2ds4 RNAi lentiviral vector / 中国实验血液学杂志
Journal of Experimental Hematology ; (6): 663-666, 2008.
Artigo em Chinês | WPRIM | ID: wpr-267915
ABSTRACT
This study was aimed to construct a lentiviral vector of RNA interfered (RNAi)-kir2ds4 gene. In accordance with study-confirmed effective sequence of siRNA targeting kir2ds4 gene, the complementary DNA containing both sense and antisense oligonuctide of the targeting sequence was designed, synthesized and inserted into pSicoR-GFP vector containing U6 promoter and GFP sequence. The resulting lentiviral vector containing kir2ds4 shRNA was named as LV-sh kir2ds4, and confirmed by PCR and sequencing. 293T cells were co-transfected with lentiviral vector LV-sh kir2ds4 and packaging system. All virus stocks were produced by Lipofectamine 2000-mediated transfection. The titer of virus was tested according to the expression level of GFP. As a result, PCR and DNA sequencing demonstrated that the lentivirus RNAi vector of kir2ds4 was constructed successfully. The titer of virus tested by expression level of GFP was 6 x 10(8) TU/ml. It is concluded that the lentivirus RNAi vector of kir2ds4 has been successfully constructed.
Assuntos
Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Dados de Sequência Molecular / Sequência de Bases / Lentivirus / RNA Interferente Pequeno / Interferência de RNA / Proteínas de Fluorescência Verde / Receptores KIR / Vetores Genéticos / Genética / Metabolismo Limite: Humanos Idioma: Chinês Revista: Journal of Experimental Hematology Ano de publicação: 2008 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Dados de Sequência Molecular / Sequência de Bases / Lentivirus / RNA Interferente Pequeno / Interferência de RNA / Proteínas de Fluorescência Verde / Receptores KIR / Vetores Genéticos / Genética / Metabolismo Limite: Humanos Idioma: Chinês Revista: Journal of Experimental Hematology Ano de publicação: 2008 Tipo de documento: Artigo