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Core-binding factor alpha1 gene recombinant virus for specifically promoting cartilage differentiation, preparation method and application thereof

A core binding factor, gene recombination technology, applied in biochemical equipment and methods, botanical equipment and methods, applications, etc., can solve the problems of chondrocyte maturation, that is, inhibition of hypertrophy, and achieve the effect of low cost and simple preparation method

Inactive Publication Date: 2011-02-23
ARMY MEDICAL UNIV
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Problems solved by technology

Although chondrocyte differentiation and chondrogenesis were normal in Cbfa1- / - mice, chondrocyte maturation, i.e. hypertrophy, was severely inhibited

Method used

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  • Core-binding factor alpha1 gene recombinant virus for specifically promoting cartilage differentiation, preparation method and application thereof
  • Core-binding factor alpha1 gene recombinant virus for specifically promoting cartilage differentiation, preparation method and application thereof
  • Core-binding factor alpha1 gene recombinant virus for specifically promoting cartilage differentiation, preparation method and application thereof

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Embodiment Construction

[0027] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the accompanying drawings. The experimental method that does not indicate specific conditions in the preferred embodiment is usually according to conventional conditions, such as described in the Molecular Cloning Experiment Guide (Third Edition, J. Sambrook et al., translated by Huang Peitang, etc., Science Press, 2002) conditions, or as recommended by the manufacturer.

[0028] 1. Preparation and identification of Cbfa1 gene recombinant adenovirus regulated by Sox9 gene promoter

[0029] 1. Preparation of Cbfa1 gene recombinant adenovirus regulated by Sox9 gene promoter

[0030] The schematic diagram of the construction of recombinant adenovirus Sox9pro-Cbfa1 is as follows figure 1 Shown, its preparation method comprises the following steps:

[0031] a. Cloning of full-length cDNA of Cbfa1 gene and construction of recombinant adenovirus shuttle plasmid of Cbfa1...

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Abstract

The invention discloses a core-binding factor alpha1 (Cbfa1) gene recombinant virus for specially promoting cartilage differentiation, a preparation method and application thereof. The recombinant virus contains one Cbfa1 gene expression box regulated and controlled by a Sox9 gene promoter; the preparation method comprises the following steps of clone of Cbfa1 gene full-length cDNA and construction of Cbfa1 gene recombinant adenovirus shuttle plasmid, clone of Sox9 gene promoter fragment and construction of the Sox9 gene promoter recombinant expression carrier, construction of Cbfa1 gene recombinant adenovirus carrier regulated and controlled by the Sox9 gene promoter, package, amplification and purification of the Cbfa1 gene recombinant adenovirus regulated and controlled by the Sox9 gene promoter and the like. The recombinant virus can be used for preparing seed cells for directionally inducing chondroblast differentiation and can efficiently and specifically ensure that the mesenchymal stem cells differentiate into chondroblast on the premise of differentiating into osteogenesis / cartilage cells.

Description

technical field [0001] The invention relates to a gene recombination virus, in particular to a core binding factor a1 (Cbfa1) gene recombination virus which specifically promotes cartilage differentiation, and also relates to a preparation method and application of the recombination virus. Background technique [0002] With the continuous deepening of histology, cytology and material science research on the process of cartilage formation, the understanding of cytokines and signaling molecules, and the interaction of cell-material interface has been continuously expanded. Some scholars have used tissue engineering technology to form tissue engineering in vitro. Cartilage, and shows the broad application prospects of tissue engineering cartilage. Although the research on tissue engineered cartilage was carried out earlier and the technology is relatively mature, the effective and reasonable source of seed cells has become a bottleneck limiting its clinical application. Among ...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N5/10C12N15/12C12N7/01C12N15/861
Inventor 杨波董世武郭洪峰康菲应大君
Owner ARMY MEDICAL UNIV
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