Construction and characterization of GFP-mfgl 2 fusion protein expression plasmid
- VernacularTitle:绿色荧光蛋白-小鼠纤维介素蛋白融合基因表达载体的构建和表达
- Author:
Zhimo WANG
;
Qin NING
- Publication Type:Journal Article
- Keywords:
Fibrinogen-like protein 2;
Green fluorescent protein;
Fusion protein
- From:
Journal of Medical Postgraduates
2003;0(04):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct and characterize GFP-mfgl 2 fusion protein expression plasmid (pEGFP-mfgl 2) and provide a direct and simplified methodology for primary assessment of the effect of mfgl 2 siRNA on the mfgl 2 gene expression. Methods: mfgl 2 cDNA was amplified from the mfgl 2 cDNA library pBluescript-m166 (pm166) of mouse genomic P1 plasmid and recloned into pEGFP-N2 upstream of GFP gene. The pEGFP-mfgl 2 was analyzed by restriction endonucleases BamH I and Hind III to ensure the orientation and the sequence. This fusion plasmid was then transfected into CHO cells and the fusion protein expression was observed by fluorescent microscope. Rusults: A 1.3 kb long cDNA was obtained. Restriction endonucleases and sequencing assays showed the correct orientation and sequence. After 24-48 hours transfection in CHO cells, the expression of pEGFP-mfgl 2 can be visualized through fluorescent microscope. Conclusion: pEGFP mfgl 2 has been constructed successfully. The recombinant vector can express GFP-mfgl 2 fusion protein. It provides a direct and simplified methodology for primary assessment of the effect of mfgl 2 siRNA on the mfgl 2 gene expression.
