Construction and identification of eukaryotic expression vector of rat Delta1 gene / 中国组织工程研究
Chinese Journal of Tissue Engineering Research
;
(53): 3331-3334, 2010.
Article
in Chinese
| WPRIM
| ID: wpr-402511
ABSTRACT
BACKGROUND:
Recent data suggested that Notch signal pathway plays important regulatory effects in peripheral transplantation immunological response, promotes differentiation of regulatory T cells, induces antigen specific immune tolerance. We proposed that Notch/Notch ligand may play important roles in MHC/TCR interface.OBJECTIVE:
To construct the eukaryotic expression vector of rat Deltai gene (Notch ligand), and to examine its expression in dendritic cells.METHODS:
The complete encoding cDNA of rat-Delta1 was isolated from bone marrow cells by reverse transcription-polymerase chain reaction (RT-PCR) and this gene was recombined into pcDNA3.1(+) plasmid vector.pcDNA3.1/Delta1 plasmid was transfected into rat dendritic cells with lipofectamine gene transfection method.RESULTS ANDCONCLUSION:
Double enzyme digestion detection demonstrated that Delta 1 had been successfully constructed in Hindilll and xbal of pcDNA3.1. A positive clone pcDNA3.1/Delta1 was delivered to Shanghai Sangon Biological Engineering Technology & Services Co., Ltd. for sequencing. Sequencing results were identical to Delta1 gene sequence in Genebank, with correct reading frame. The Delta 1 gene-transfected dendritic cells showed similar morphology as their parent cells. Western blotting assay detected that Delta 1 expression was significantly increased in cells. The eukaryotic expression vector pcDNA3.1/Delta1 was constructed, and significant increase of Delta 1 expression was detected after transfection.
Full text:
Available
Index:
WPRIM (Western Pacific)
Type of study:
Diagnostic study
Language:
Chinese
Journal:
Chinese Journal of Tissue Engineering Research
Year:
2010
Type:
Article
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