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Building method of zebra fish intervertebral disc injury model

A damage model and construction method technology, applied in the field of genetic engineering, can solve the problems of impractical real-time dynamic labeling of cell migration, differentiation process, general representation, and high economic cost

Active Publication Date: 2017-06-20
STOMATOLOGICAL HOSPITAL OF CHONGQING MEDICAL UNIV
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0003] At present, it is more common for the study of spinal degenerative diseases to establish an animal model of intervertebral disc degeneration by surgically injuring the lumbar intervertebral disc in mice, but the defects of this modeling are very obvious, such as large surgical trauma, The economic cost is high, and the mouse embryo develops in the mother mouse, which is not conducive to the study of the early development and progress of the spine, and is not suitable for the migration and differentiation process of real-time dynamic marker cells in vivo, so we think that the mouse is the preferred mammalian animal model representative of general

Method used

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  • Building method of zebra fish intervertebral disc injury model
  • Building method of zebra fish intervertebral disc injury model
  • Building method of zebra fish intervertebral disc injury model

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Experimental program
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Embodiment 1

[0031] The construction method of the construction method of the zebrafish intervertebral disc injury model of the present invention, its operation method is:

[0032] Step 1. Construction of pBluescriptII-KS-twhh-Dendra2-NTR recombinant plasmid

[0033] 1. Experimental materials

[0034] 1.1 Experimental animals

[0035] The experimental research was carried out in the cultivation base of the State Key Laboratory of Ecological Environment and Biological Resources in the Three Gorges Reservoir Area, Chongqing. Wild-type zebrafish (AB line) were bred and raised under standard experimental conditions in accordance with the regulations of the Experimental Animal Management Committee.

[0036] 1.2 Experimental reagents

[0037] The PCR primer synthesis and sequence determination in this experiment were operated by Shanghai Handsome Biotechnology Co., Ltd.;

[0038] Primer STAR DNA polymerase was from TaKaRa, Japan;

[0039] LA Taq DNA Polymerase was from TaKaRa, Japan;

[004...

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Abstract

The invention discloses a building method of a zebra fish intervertebral disc injury model. The method comprises DNA (deoxyribonucleic acid) polymerase chain type reaction, and is characterized in that the primers and the sequences used in the DNA polymerase chain type reaction as shown as follows: a positive primer: 5'-GTCGACGTACTGTATCTGTGCTGATTCAC-3'; a reverse primer: 5'-GACCAGAAGAGATAATTCAAACGT-3'. The building method aims at solving the technical problems to provide the building method of the zebra fish intervertebral disc injury model for providing the theoretical basis for the intervertebral disc degeneration study.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to a method for constructing a zebrafish intervertebral disc injury model. Background technique [0002] Intervertebral disc degeneration caused by the accumulation of pressure and wear with age is a unique aging disease of the spine, which can cause 20% of low back pain (LBP). Intervertebral disc degeneration is the main cause of chronic low back pain. cause. The development, degeneration, regeneration and repair of intervertebral disc is a complicated process, the exact molecular biological mechanism is not yet clear, and many factors are involved, which needs to be further studied. Intervertebral disc herniation, low back pain, and cervical spondylosis are common and frequently-occurring diseases in clinic, and their pathogenesis is directly related to intervertebral disc degeneration. Studying the degeneration mechanism and repair mechanism of the intervertebral disc can pro...

Claims

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Application Information

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IPC IPC(8): C12N15/85A01K67/027
CPCA01K67/0278A01K2207/15A01K2217/05A01K2227/40A01K2267/03C12N15/8509
Inventor 杨德琴黄兴罗凌飞杨其芬黄文明
Owner STOMATOLOGICAL HOSPITAL OF CHONGQING MEDICAL UNIV
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