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msCT-acAP5 fusion protein transgenic engineering strain

A technology of transgenic engineering, msct-acap5, applied in the field of fusion protein transgenic engineering strains, can solve the problems of patient inconvenience, antagonistic reaction, etc., and achieve the effect of safe use and wide application prospects

Inactive Publication Date: 2014-07-09
河北尚墨农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

If the patient takes multiple drugs to treat osteoporosis and thrombosis at the same time, drug antagonistic reactions are likely to occur; if the time of taking drugs is to be staggered, on the one hand, the half-life of the drug must be considered, and on the other hand, the effective concentration of the drug must also be considered, and it will cause inconvenience to the patient.
[0005] At present, there is no drug that can effectively treat osteoporosis and thrombosis at the same time

Method used

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  • msCT-acAP5 fusion protein transgenic engineering strain
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  • msCT-acAP5 fusion protein transgenic engineering strain

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Experimental program
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specific Embodiment approach 1

[0017] Specific embodiment 1: In this embodiment, the msCT-AcAP5 fusion protein transgenic engineering strain is named Escherichia coli BL21(DE3)-msCT / AcAP5, and Escherichia coli BL21(DE3)-msCT / AcAP5 was prepared according to the following steps:

[0018] 1. Digest the plasmid vector pUC-msCT / AcAP5 containing the msCT-AcAP5 fusion protein gene with BamH I and EcoR I to obtain the DNA fragment of the msCT-AcAP5 fusion protein;

[0019] 2. Digest plasmid pGEX-6P-1 with BamHI and EcoRI;

[0020] 3. The DNA fragment of the msCT-AcAP5 fusion protein was enzymatically ligated with the vector pGEX-6P-1 after double enzyme digestion, and then placed at 16°C for overnight connection to obtain the vector pGEX-msCT / AcAP5;

[0021] 4. Transform Escherichia coli BL21(DE3) with vector pGEX-msCT / AcAP5, and select positive recombinants to obtain msCT-AcAP5 fusion protein transgenic engineering bacteria Escherichia coli BL21(DE3)-msCT / AcAP5.

[0022] Step 4 of this embodiment transforms Esche...

specific Embodiment approach 2

[0023] Specific embodiment two: the difference between this embodiment and specific embodiment one is that: in step one, the enzyme digestion reaction system of plasmid vector pUC-msCT / AcAP5 is

[0024]

[0025]

[0026] Other steps and parameters are the same as those in the first embodiment.

[0027] In the first step of this embodiment, the enzyme digestion reaction was carried out at 37°C for 10 hours, followed by 1.5% agarose gel electrophoresis, and the target fragment was purified and recovered with DNA GEL EXTRACTION KIT.

specific Embodiment approach 3

[0028] Specific embodiment three: the difference between this embodiment and specific embodiment one or two is: the enzyme digestion reaction system of plasmid pGEX-6P-1 in step two is

[0029]

[0030] Other steps and parameters are the same as those in Embodiment 1 or 2.

[0031] In the second step of this embodiment, the enzyme digestion reaction was carried out at 37°C for 10 hours, followed by 0.6% agarose gel electrophoresis, and the target fragment was purified and recovered with DNA GEL EXTRACTION KIT.

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Abstract

The invention provides an msCT-AcAP5 fusion protein transgenic engineering strain, relating to a fusion protein transgenic engineering strain and overcoming the defect that drugs for effectively treating osteoporosis and thrombus at the same time do not exist at present. The msCT-AcAP5 fusion protein transgenic engineering strain escherichia coli BL21 (DE3-msCT / AcAP5) is prepared by the following steps of: 1. obtaining a DNA (deoxyribonucleic acid) segment of the fusion protein; 2. carrying out double digestion on a plasmid; 3. carrying out enzyme connection; and 4. transforming the escherichia coli BL21. The strain can be used in the field of drug preparation.

Description

technical field [0001] The invention relates to a fusion protein transgenic engineering strain. Background technique [0002] Thrombosis is a multifactorial process involving many interacting genetic and environmental factors. However, the coagulation system of the elderly has its particularity. The increase of fibrinogen (FIB) content, tissue plasminogen activator and plasminogen activator inhibitor complex in the elderly all lead to Hypercoagulable state, so it is easier to form blood clots; Among them, studies have found that the incidence of blood clots in postmenopausal women is much higher than that in men. [0003] The incidence of osteoporosis in the elderly is high. There are 200 million osteoporosis patients in the world, and there are more women than men. According to the standards of the World Health Organization (WHO), the results of the US National Health and Nutrition Survey (NHANESⅢ, 1988-1994) showed that osteoporosis seriously affects the quality of life ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/21C12N15/70C12R1/19
Inventor 余琼
Owner 河北尚墨农业科技有限公司