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Preparation method of antivirus and inactivated yeast engineered strains

A technology for inactivating yeast and antiviral activity, applied in the biological field, can solve problems such as difficult poultry breeding

Inactive Publication Date: 2017-12-29
GUIZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, genetically modified active engineering bacteria are subject to the constraints of relevant transgenic safety management regulations in countries around the world, including my country, due to possible hidden dangers such as potential ecological safety, so it is difficult to directly apply them to poultry farming

Method used

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  • Preparation method of antivirus and inactivated yeast engineered strains
  • Preparation method of antivirus and inactivated yeast engineered strains
  • Preparation method of antivirus and inactivated yeast engineered strains

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The construction of the preparation method of embodiment 1 antiviral inactivated yeast engineered bacteria

[0027] 1) strain

[0028] INVSC1 / pYEIFNG and INVSC1 / pYEIFNR recombinant Saccharomyces cerevisiae contain ChIFN-γ (U27465.1) and ChIFN-α genes (U07868.1) respectively, which were created by the Agricultural Bioengineering Research Institute of Guizhou University and stored in a -80°C refrigerator.

[0029] 2) Screening, cultivation and identification of engineering strains

[0030] 2.1) Use the SC selection medium to screen the engineering bacteria, inoculate the engineering strains on the SC solid selection medium, culture them at 30°C until a single colony is formed, and store them at -80°C for later use;

[0031] 2.2) Take INVSC1 / pYEIFNG and INVSC1 / pYEIFNR engineered bacteria that were stored at -80°C or cultured immediately and spread them on YPD solid medium, and culture them at 30°C for 24-48 hours; pick a single colony of activated engineered bacteria and ...

Embodiment 2

[0052] The inhibitory action of embodiment 2 crude protein extracts to NDV

[0053] Chicken embryo fibroblasts were 5.0×10 4 Inoculate in a 96-well plate at a density of 1 / ml, inoculate 100 μL in each well, and store at 37°C in 5% CO 2 Cultivate in an incubator until 80% of the cells adhere to the wall, and inoculate 100 μL of the mixture in each well (the composition of the mixture is 50 μL of DMEM with 10% FBS and 50 μL of lysate in a 1:1 ratio). After culturing for 10 h, the liquid in the 96-well plate was discarded, and 100 TCID 50 Inoculate 100 μL of the virus solution into each well, place it in the incubator for 2 hours, discard the virus solution and wash it twice with Hanks solution, inoculate the new maintenance solution, and continue to cultivate for 48 hours, add 20 μL of 2 mg / ml MTT solution to each well, and continue to cultivate for 4 hours. After the incubation, the liquid in the wells was sucked out, and DMSO solution was added at 100 μL / well, and the crysta...

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Abstract

The invention discloses a preparation method of antivirus and inactivated yeast engineered strains. The preparation method includes that the engineered strains subjected to induced expression of recombinant chicken interferon are subjected to complete inactivation treatment for 30 minutes at 71DEG C, and the inactivated engineered strains are confirmed to contain rChIFN-alpha and rChIFN-gamma have inhibiting effect on Newcastle disease virus by the aid of MTT experiments which indicate that the inactivated engineered strains with ecological safety still keeps antivirus biological activities. The preparation method is simple and easy to implement and low in cost, and obvious function on virus disease control of poultry is achieved.

Description

technical field [0001] The application belongs to the field of biotechnology, and in particular relates to a preparation method of an antiviral inactivated yeast engineered bacterium. Background technique [0002] Saccharomyces cerevisiae is a green and safe probiotic feed additive, which plays an important role in enhancing the immunity of animals and improving the balance of intestinal microorganisms. Chicken interferon (ChIFN) is a kind of animal cytokine with high efficiency and broad-spectrum antiviral effect. important drug for the disease. [0003] At present, the production of poultry interferon at home and abroad is mainly produced by genetic engineering technology and purified into injection solution, which has defects such as high production cost and prone to stress, which limits the wide application of poultry interferon in the breeding industry. application. It is of great significance to study the poultry interferon transgenic probiotic engineering bacteria ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/19C12Q1/68C12Q1/04A23K10/16A23K20/147G01N33/68C12R1/865
CPCA23K10/16C07K14/465C12Q1/686G01N33/6866C12Q2531/113
Inventor 宋莉张冬生赵德刚
Owner GUIZHOU UNIV
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