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A kind of construction method of deer antler bioreactor

A bioreactor, construction method technology, applied in biochemical equipment and methods, botanical equipment and methods, cells modified by introducing foreign genetic material, etc. , the effect of increasing production

Inactive Publication Date: 2018-06-15
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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  • A kind of construction method of deer antler bioreactor
  • A kind of construction method of deer antler bioreactor
  • A kind of construction method of deer antler 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 0.01% poly-lysine to each plate, reverse left and right, spread the liquid on the bottom of the plate, incubate at room temperature for 10min to remove poly-lysine, add 13mL 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, use 1.5mL serum-free Opti-MEM to dilute 24μg DNA (the ratio of pLVTHM:pCMV-dr8.91:pMD2.G plasmid mass is 2:1.5:0.75), use 1.5mL serum-free Opti-MEM to dilute 51μL Transfection reagent Lipofectamine2000, incubated at room temperature for 5min. The diluted DNA and diluted LIPOFECTAMINE 2000 were mixed together, and the mixture was incubated at room temperature for 30 minutes. Aspirate the culture medium in the 10cm cell culture dish, add the complex to the cell culture dish, shake the culture plate, and mix ge...

Embodiment 2

[0055] 1. Preparation and purification of lentiviral particles

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

Embodiment 3

[0066] 1. Preparation and purification of lentiviral particles

[0067] The viral plasmid pLVTHM was transformed, and the galactosidase gene (LacZ) was inserted, and the transformed plasmid was named pLVTHM-LacZ, and the endotoxin-free large-scale extraction was carried out for future use. Take out a certain number of 10cm cell culture dishes, add 4mL 0.01% poly-lysine to each plate, reverse left and right, spread the liquid on the bottom of the plate, incubate at room temperature for 10min to remove poly-lysine, add 13mL 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, use 1.5mL serum-free Opti-MEM to dilute 24μg DNA (the ratio of pLVTHM-LacZ:pCMV-dr8.91:pMD2.G plasmid mass is 2:1.5:0.75), use 1.5mL serum-free Opti-MEM Dilute 51 μL of transfection reagent Lipofectamine 2000 and incubate at room temperature for 5 min. The diluted DNA and diluted LIPOFECTAMINE...

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Abstract

The invention discloses a method for constructing a deer antler bioreactor, and belongs to the technical field of genetic engineering. The method provided by the present invention is to prepare lentiviral particles containing exogenous genes, combine lentiviral particles with magnetic carriers, infect the periosteum of antler-growing areas, detect the expression of marker genes or exogenous genes, and obtain antler bioreactors. The present invention can artificially transform beneficial components in velvet antler, increase the content of cartilage components in velvet antler, increase the output of velvet antler wax flakes, increase the added value of velvet antler, and increase the income of farmers; it can also increase the content of certain beneficial components in velvet antler; the velvet antler produced by the invention The bioreactor can be regenerated periodically with the antler every year, and can be obtained repeatedly, that is, one transformation is valid for life; the transformation of the deer individual in the present invention is limited to the periosteum tissue that determines the production of antler, and the exogenous gene is only expressed in the epigenetic antler tissue. More in line with animal ethics requirements. The invention is suitable for constructing antler bioreactors.

Description

technical field [0001] The invention relates to a construction method of a velvet antler bioreactor, belonging to the technical field of genetic engineering. Background technique [0002] In recent years, due to the achievements in the field of molecular biology research, the vigorous development of transgenic animal bioreactors has been promoted. At present, the bioreactors that have been successfully researched and can be industrialized mainly include bacterial reactors, insect baculovirus systems, fungal systems, mammalian cells, and living bioreactors. The existing bioreactors have their own characteristics. For example, the bacterial reactor has a large expression amount and low cost, but many mammalian gene products expressed are not biologically active; the expression system of mammalian cells is extremely expensive and the expression amount Very low, but can express those mammalian active factors that low expression systems cannot express. Once established, the liv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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