Construction and characterization of a recombinant human adenovirus vector expressing bone morphogenetic protein 2

Zhang,Zheng; Wang,Guoxian; Li,Chen; Liu,Danping

doi:10.3892/etm.2013.1162

August 2013 Volume 6 Issue 2

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August 2013 Volume 6 Issue 2

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Article

Construction and characterization of a recombinant human adenovirus vector expressing bone morphogenetic protein 2

Authors:
- Zheng Zhang
- Guoxian Wang
- Chen Li
- Danping Liu
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Affiliations: Department of Orthopaedics, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning 121001, P.R. China, Department of Pharmacology, Liaoning Medical University, Jinzhou, Liaoning 121000, P.R. China, Biological Samples Library, The First Affiliated Hospital of Liaoning Medical University, Jinzhou, Liaoning 121001, P.R. China
Pages: 329-334
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Published online on: June 17, 2013

https://doi.org/10.3892/etm.2013.1162
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Abstract

The aim of this study was to construct and characterize a novel recombinant human adenovirus vector expressing bone morphogenetic protein 2 (BMP2) and green fluorescent protein (GFP). The BMP2 gene in the plasmid pcDNA3-BMP2 was sequenced and the restriction enzyme recognition sites were analyzed. Following mutagenesis using polymerase chain reaction (PCR), the gene sequence after the translation termination codon was removed and new restriction sites were added. The mutated BMP2 gene (BMP2+ gene) was cloned into an adenovirus shuttle vector to obtain pShuttle cytomegalovirus (CMV)-BMP2+-internal ribosome entry site (IRES)-hrGFP-1. The adenovirus plasmid pAd CMV-BMP2+-IRES-hrGFP-1 was constructed by homologous recombination and was transfected into HEK293A cells, followed by adenovirus packaging. pAd CMV-BMP2 was used as the control. The two types of adenovirus were transfected into marrow stromal cells (MSCs). The expression of BMP2 and GFP, as well as the alkaline phosphatase (ALP) activity of expressed BMP2 were detected. Following mutagenesis, the BMP2 gene sequence and recombinant adenovirus vector were as predicted. The novel adenovirus vector expressed both BMP2 and GFP, indicating that a novel recombinant human adenovirus vector expressing BMP2 had been successfully constructed.

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Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Construction and characterization of a recombinant human adenovirus vector expressing bone morphogenetic protein 2

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