Expression vector for the inhibitor of growth-1 gene is constructed and the NLS-GFP fusion protein expresses in MRC-5 cells / 中华医学遗传学杂志
Chinese Journal of Medical Genetics
;
(6): 330-332, 2006.
Artigo
em Chinês
| WPRIM
| ID: wpr-263783
ABSTRACT
<p><b>OBJECTIVE</b>To construct the NLS(ING1)-GFP vector, transfer it into MRC-5 cells and establish a cell model expressing NLS (ING1)-GFP fusion protein.</p><p><b>METHODS</b>Firstly, cDNA fragment of nuclear locating sequence (NLS) of inhibitor of growth-1 gene (ING1) was gained by RT-PCR and inserted into multi-clone site of pEGFP-C1 to construct the NLS (ING1)-GFP expression vector. Then the vector was used to transfect the MRC-5 cells to observe the subcellular signal localization of green fluorescence protein (GFP).</p><p><b>RESULTS</b>We successfully constructed the expressing vector of NLS (ING1)-GFP fusion protein. After transferring the fusion expressing vector into MRC-5 cells, we observed that green fluorescence signal located in the cell nucleus. However, the green fluorescence signal located in the cytoplasm in MRC-5 cells transfected with pEGFP-C1 control only expressing GFP.</p><p><b>CONCLUSION</b>In living cells, physiologically p33 ING1b locates absolutely in nucleus. The p33(ING1b) NLS plays a decisive role in the transporting process of subcellular localization.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Proteínas Recombinantes de Fusão
/
Dados de Sequência Molecular
/
Proteínas Nucleares
/
Sequência de Bases
/
Transfecção
/
Linhagem Celular
/
Análise de Sequência de DNA
/
Reação em Cadeia da Polimerase Via Transcriptase Reversa
/
Proteínas Supressoras de Tumor
/
Peptídeos e Proteínas de Sinalização Intracelular
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Medical Genetics
Ano de publicação:
2006
Tipo de documento:
Artigo
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