Construction of human bone morphogenetic protein 2 and histidine eukaryotic expression plasmid and synthesis of chitosan/pIRES2-EGFP-hBMP2-His nanoparticles / 华西口腔医学杂志
West China Journal of Stomatology
;
(6): 493-497, 2014.
Article
in Chinese
| WPRIM
| ID: wpr-231818
ABSTRACT
<p><b>OBJECTIVE</b>To clone and construct a eukaryotic expression vector of human bone morphogenetic protein (BMP) 2 and histidine in vitro and synthesize chitosan (CS)/pIRES2-EGFP-hBMP2-His nanoparticles.</p><p><b>METHODS</b>pMD18T-hBMP2-His was digested by EcoR I and BamH I to obtain the hBMP2-His gene, which was inserted into pIRES2-EGFP to form pIRES2-EGFP-hBMP2-His. Afterward, CS, which exhibited five different molecular weights and deacetylation degrees, was complexed with pIRES2-EGFP-hBMP2-His to form CS/pIRES2-EGFP-hBMP2-His nanoparticles; in this procedure, a desolvent method was used at different N/P ratios (amino in CS to phospho in plasmid DNA). The gene-encapsulating ability of CS was evaluated by agarose gel electrophoresis and fluorescence spectrophotometry; size, distribution, and potential were analyzed using a ZetaPALS analyzer. The shape of the nanoparticles was observed under an atomic force microscope.</p><p><b>RESULTS</b>1) pIRES2-EGFP-hBMP2-His was constructed after the cloned hBMP2-His gene was confirmed by sequencing. 2) CS/pIRES2-EGFP-hBMP2-His nanoparticles were synthesized and pIRES2-EGFP-hBMP2-His was packaged by CS. 3) CS/pIRES2-EGFP-hBMP2-His nanoparticles were globular with an average size of 111.7 nm to 3,214.2 nm and an average zeta-potential of 4.93 mV to 16.79 mV.</p><p><b>CONCLUSION</b>CS/pIRES2-EGFP-hBMP2-His nanospheres are successfully synthesized.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
Chitosan
/
Green Fluorescent Proteins
/
Nanoparticles
/
Bone Morphogenetic Protein 2
/
Genetic Vectors
/
Histidine
Limits:
Humans
Language:
Chinese
Journal:
West China Journal of Stomatology
Year:
2014
Type:
Article
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