Expression of VEGF165 and VEGF121 genes in MC3T3-E1 cell line and biological function of their products / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
;
(6): 281-286, 2006.
Artigo
em Chinês
| WPRIM
| ID: wpr-332158
ABSTRACT
<p><b>OBJECTIVE</b>To construct vectors stably expressing bioactive VEGF165 and VEGF121 in MC3T3-E1 cells.</p><p><b>METHODS</b>VEGF165 and VEGF121 genes were obtained by RT-PCR from HL-60 cells and inserted into eukaryotic expression plasmid pcDNA3.1(+). The constructed plasmids containing right sequence of target genes were transfected into MC3T3-E1 cells by lipofectin. The cells were limitedly diluted twice and selected by G418, the expression of VEGF was tested by ELISA, Western-blot and immunofluorescence essay. The bioactivity of expressed products was proved by MTT method.</p><p><b>RESULT</b>DNA sequencing indicated the sequences of the obtained VEGF165 and VEGF121 were identical to the those reported in Genebank, and the newly-constructed plasmids were VEGF165/pcDNA3.1(+) and VEGF121/pcDNA3.1(+). The results of ELISA, Western-blot and immunofluorescence showed that the transfected cells could express VEGF165 and VEGF121 proteins. And MTT proved the proteins were bioactive.</p><p><b>CONCLUSION</b>The transfected MC3T3-E1 cells could stably express high levels of bioactive VEGF165 and VEGF121.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Plasmídeos
/
Proteínas Recombinantes
/
Transfecção
/
Células 3T3
/
Células HL-60
/
Fator A de Crescimento do Endotélio Vascular
/
Genética
Limite:
Animais
/
Humanos
Idioma:
Chinês
Revista:
Journal of Zhejiang University. Medical sciences
Ano de publicação:
2006
Tipo de documento:
Artigo
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