Han Qian-qian, Ge Shao-hua, Yan Xiang-zhen, Yang Pi-shan
Department of Periodontology, School of Stomatology, Shandong University, Key Lab of Oral Biomedicine of Shandong Province, Jinan 250012, Shandong Province, China.
Shanghai Kou Qiang Yi Xue. 2013 Feb;22(1):25-9.
To construct and confirm a recombinant lentiviral vector containing human bone morphogenetic protein 2 (hBMP2) and human nerve growth factor (hNGF).
The Neomycin gene was digested from pLentiTrident1-EGFP-Neo and then was subcloned into lentiviral vector. The hBMP2 and hNGF genes were amplified by polymerase chain reaction (PCR), and then the PCR product was inserted to proper sites of the vector. Finally, the recombinant vector pLentiTrident1-hBMP2-Neo-hNGF was confirmed by agarose gel electrophoresis and DNA sequence analysis.
The construction of recombinant lentiviral vector pLentiTrident-hBMP2 -Neo-hNGF was confirmed through restriction enzyme maping analysis and DNA sequencing.
The recombinant lentiviral vector which can coexpress hBMP2 and hNGF is successfully constructed,which lays a solid foundation of studying the effect of neuro factors on bone regeneration.
构建并鉴定一种包含人骨形态发生蛋白2（hBMP2）和人神经生长因子（hNGF）的重组慢病毒载体。
从pLentiTrident1-EGFP-Neo中酶切出新霉素基因，然后亚克隆至慢病毒载体。通过聚合酶链反应（PCR）扩增hBMP2和hNGF基因，随后将PCR产物插入载体的合适位点。最后，通过琼脂糖凝胶电泳和DNA序列分析鉴定重组载体pLentiTrident1-hBMP2-Neo-hNGF。
通过限制性酶切图谱分析和DNA测序证实了重组慢病毒载体pLentiTrident-hBMP2-Neo-hNGF的构建。
成功构建了可共表达hBMP2和hNGF的重组慢病毒载体，为研究神经因子对骨再生的作用奠定了坚实基础。
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