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Construction method for cornua cervi pantotrichum bioreactor

A technology of bioreactor and construction method, which is applied in biochemical equipment and methods, botanical equipment and methods, cells modified by introducing foreign genetic material, etc., can solve the problems of breast reactor development period limitation, etc. , the effect of increasing production

Inactive Publication Date: 2015-07-15
INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, mammary gland reactors are not perfect. For example, the development cycle of mammary gland reactors is limited by the growth and reproduction cycle of animals.
In addition, not all bioactive factors are suitable for breast reactor expression

Method used

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  • Construction method for cornua cervi pantotrichum bioreactor
  • Construction method for cornua cervi pantotrichum bioreactor
  • Construction method for cornua cervi pantotrichum bioreactor

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] 1. Preparation and purification of lentiviral particles

[0045] Take out a certain number of 10cm cell culture dishes, add 4mL of 0.01% poly-lysine to each plate, invert left and right to make the liquid spread on the bottom of the plate, incubate at room temperature for 10min to remove poly-lysine, add 13mL of DMEM to each dish Culture medium (containing 10% FBS) and a certain number of 293t cells were cultured in a cell incubator (37°C, 5% CO 2 ). For each plate of cells, dilute 24 μg DNA with 1.5 mL of serum-free Opti-MEM (pLVTHM:pCMV-dr8.91:pMD2.G plasmid mass ratio is 2:1.5:0.75) and 51 μL with 1.5 mL of serum-free Opti-MEM Transfection reagent Lipofectamine2000, incubated at room temperature for 5min. The diluted DNA and the diluted LIPOFECTAMINE 2000 were mixed together, and the mixture was incubated at room temperature for 30 min. Aspirate the culture medium in the 10cm cell culture dish, add the complex to the cell culture dish, shake the culture plate, and...

Embodiment 2

[0055] 1. Preparation and purification of lentiviral particles

[0056] Take out a certain number of 10cm cell culture dishes, add 4mL of 0.01% poly-lysine to each plate, invert left and right to make the liquid spread on the bottom of the plate, incubate at room temperature for 10min to remove poly-lysine, add 13mL of DMEM to each dish Culture medium (containing 10% FBS) and a certain number of 293t cells were cultured in a cell incubator (37°C, 5% CO 2 ). For each plate of cells, dilute 24 μg DNA with 1.5 mL of serum-free Opti-MEM (pLVTHM:pCMV-dr8.91:pMD2.G plasmid mass ratio is 2:1.5:0.75) and 51 μL with 1.5 mL of serum-free Opti-MEM Transfection reagent Lipofectamine2000, incubated at room temperature for 5min. The diluted DNA and the diluted LIPOFECTAMINE 2000 were mixed together, and the mixture was incubated at room temperature for 30 min. Aspirate the culture medium in the 10cm cell culture dish, add the complex to the cell culture dish, shake the culture plate, and...

Embodiment 3

[0066] 1. Preparation and purification of lentiviral particles

[0067] The virus plasmid pLVTHM was transformed, and the galactosidase gene (LacZ) was inserted, and the successfully transformed plasmid was named pLVTHM-LacZ, and the endotoxin-free large-scale extraction was carried out and reserved for future use. Take out a certain number of 10cm cell culture dishes, add 4mL of 0.01% poly-lysine to each plate, invert left and right to make the liquid spread on the bottom of the plate, incubate for 10min at room temperature to remove poly-lysine, add 13mL of DMEM to each dish Culture medium (containing 10% FBS) and a certain number of 293t cells were cultured in a cell incubator (37°C, 5% CO 2 ). For each plate of cells, dilute 24 μg DNA in 1.5 mL of serum-free Opti-MEM (pLVTHM-LacZ:pCMV-dr8.91:pMD2.G plasmid mass ratio is 2:1.5:0.75), use 1.5 mL of serum-free Opti-MEM Dilute 51 μL of transfection reagent Lipofectamine 2000 and incubate at room temperature for 5 min. The d...

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Abstract

The invention discloses a construction method for a cornua cervi pantotrichum bioreactor, and belongs to the technical field of genetic engineering. The construction method comprises the following steps: preparing slow virus particles containing exogenous genes, infecting antlerogenic periosteum after combining the slow virus particles with a magnetic carrier, detecting expression conditions of marker genes or exogenous genes to obtain the cornua cervi pantotrichum bioreactor. According to the construction method, beneficial components in the cornua cervi pantotrichum can be artificially modified, the content of cartilage components in the cornua cervi pantotrichum is increased, the yield of cornua cervi pantotrichum wax sheets is increased, the additional value of the cornua cervi pantotrichum is increased to increase the income of households, and the content of certain beneficial component in the cornua cervi pantotrichum further can be increased. The cornua cervi pantotrichum bioreactor constructed by the construction method can be repeatedly obtained along with annular periodic regeneration of the cornua cervi pantotrichum, so that once modification is perpetually effective. The bioreactor is only restricted to determine the periosteum tissue for cornua cervi pantotrichum generation, and the exogenous genes are only expressed in epigenetic cornua cervi pantotrichum tissue, so that the animal ethic requirements are met better. The construction method is suitable for constructing the cornua cervi pantotrichum bioreactor.

Description

technical field [0001] The invention relates to a construction method of a deer antler bioreactor, belonging to the technical field of genetic engineering. Background technique [0002] In recent years, the development of bioreactors for transgenic animals has been promoted due to the achievements in the field of molecular biology research. At present, the bioreactors that have been successfully researched and can be industrially produced mainly include bacterial reactors, insect baculovirus systems, fungal systems, mammalian cells and living bioreactors. The existing bioreactors have their own characteristics. For example, bacterial reactors have a large amount of expression and low cost, but many mammalian gene products expressed have no biological activity. Very low, but can express those mammalian active factors that cannot be expressed by lower expression systems. The in vivo expression system represented by the animal mammary gland reactor, once established, not only...

Claims

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Application Information

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IPC IPC(8): C12N15/867C12N5/10
Inventor 李春义褚文辉赵海平孙红梅刘振金美伶
Owner INST OF SPECIAL ANIMAL & PLANT SCI OF CAAS
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