[Construction of sense and antisense eukaryotic expression vector of novel gene Collectrin and its function in cell growth].
Beijing Da Xue Xue Bao Yi Xue Ban
; 36(2): 181-4, 2004 Apr.
Article
en Zh
| MEDLINE
| ID: mdl-15100739
ABSTRACT
OBJECTIVE:
To construct sense and antisense eukaryotic expression vector of novel gene Collectrin and identify its function in cell growth.METHODS:
The open reading frame of Collectrin was amplified by PCR and inserted into pcDNA3.1/V5-His plasmid. The recombinant plasmid was identified by restriction enzyme analysis and sequencing analysis. The recombinant plasmid was transfected into M-1 cell by using lipofectin mediation after being identified by restriction enzyme analysis and sequencing analysis. RT-PCR and Western blot were performed to identify the expression of Collectrin. Beta-Gal staining was used to define the effect of transfection. The growth of M-1 cells was examined by MTT and cell counting.RESULTS:
Compared with control group, the expression of Collectrin was decreased significantly at both nucleotide and protein levels transfected by antisense vector, but elevated in sense group. The cell growth was blocked after being transfected by antisense plasmid.CONCLUSION:
The sense and antisense eukaryotic expression vector of novel gene Collectrin was successfully constructed. Collectrin was one of basic factors in cell growth.
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Banco de datos:
MEDLINE
Asunto principal:
Glicoproteínas de Membrana
Tipo de estudio:
Prognostic_studies
Límite:
Animals
Idioma:
Zh
Año:
2004
Tipo del documento:
Article