[Construction and expression of fusion gene eukaryotic expression plasmid of pEGFP-C(3)-insig2 and its influence to downstream genes].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
; 23(4): 309-11, 2007 Apr.
Article
em Zh
| MEDLINE
| ID: mdl-17428382
AIM: To construct the eukaryotic expression plasmid of insig2 gene and detect the expression of downstream gene adiponectin and adipocyte fatty acid-binding protein 2 (AP2) after the transfection of 3T3-L1 cells. METHODS: Insig2 gene of the mouse was amplified by RT-PCR and then cloned into the eukaryon expression vector pEGFP-C(3), After confirmed by double restriction enzyme digestion analysis and DNA sequencing, pEGFP-C(3)-insig2 was transfected into 3T3-L1 cells by lipofectamine 2000. The expression of insig2 and downstream gene in the 3T3-L1 cells were detected by RT-PCR and fluorescence microscope. RESULTS: The eukaryotic expression plasmid of pEGFP-C(3)-insig2 was constructed. The expression of fusion protein in the endochylema was confirmed. The transcription of adiponectin mRNA and AP2 mRNA was down-regulated after transfection for 24 h and 72 h. CONCLUSION: The eukaryotic expression plasmid of pEGFP-C(3)-insig2 is successfully constructed. The transfected insig2 may have an effect on fat metabolism of 3T3-L1 cells.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Plasmídeos
/
Proteínas Recombinantes de Fusão
/
Proteínas de Fluorescência Verde
/
Proteínas de Membrana
Limite:
Animals
Idioma:
Zh
Revista:
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
Assunto da revista:
ALERGIA E IMUNOLOGIA
Ano de publicação:
2007
Tipo de documento:
Article
País de afiliação:
China
País de publicação:
China