华西口腔医学杂志Issue(5)：493-497,5.DOI:10.7518/hxkq.2014.05.016
人骨形态发生蛋白2真核表达载体构建及其基因/
Construction of human bone morphogenetic protein 2 and histidine eukaryotic expression plasmid and synthesis of ;chitosan/pIRES2-EGFP-hBMP2-His nanoparticles
摘要
Abstract
Objective 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. Methods pMD18T-hBMP2-His was digested by EcoRⅠand BamHⅠto 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. Results 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. Conclusion CS/pIRES2-EGFP-hBMP2-His nanospheres are successfully syn-thesized.关键词
人骨形态发生蛋白2/真核表达载体/壳聚糖/纳米复合体Key words
human bone morphogenetic protein 2/eukaryotic expression vector/chitosan/nanoparticle分类
医药卫生引用本文复制引用
杨晓喻,李世轶,张迪,吴颖,杨涛,刘长虹..人骨形态发生蛋白2真核表达载体构建及其基因/[J].华西口腔医学杂志,2014,(5):493-497,5.基金项目
国家自然科学基金资助项目 （）
