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Preparation method of periodic Wuchereria malayi compound polyvalent protein vaccine

A Malayan worm and protein vaccine technology is applied in the directions of medical preparations, drug combinations, and pharmaceutical formulations containing active ingredients, which can solve the lack of eukaryotic gene expression regulation mechanism protein processing and modification ability, and the expression products are easy to lose immunity. activity, etc.

Inactive Publication Date: 2014-06-04
NANTONG UNIVERSITY
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AI Technical Summary

Problems solved by technology

The prokaryotic expression system is the most mature exogenous protein expression system. It is easy to operate and can express a large amount of protein. However, it lacks the gene expression regulation mechanism of eukaryotes and the ability of protein processing and modification. The expression product cannot spontaneously fold and coil to form a certain spatial structure and Proteins with specific biological functions often exist in the form of inclusion bodies, and after denaturation and renaturation, the expressed products tend to lose their immune activity

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  • Preparation method of periodic Wuchereria malayi compound polyvalent protein vaccine
  • Preparation method of periodic Wuchereria malayi compound polyvalent protein vaccine
  • Preparation method of periodic Wuchereria malayi compound polyvalent protein vaccine

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Embodiment Construction

[0133] A kind of preparation method of period type malayia filarial compound multivalent protein vaccine comprises the following steps:

[0134] (1) Extraction of periodic filaria pcDNA3.1(+)-BmCPI / BmGAPDH recombinant plasmid:

[0135] (1) After thawing the strain containing the recombinant plasmid at room temperature, take 50ul and add it to 100ml LB solution containing 50ul and 100mg / ml ampicillin sodium, place it on a shaker at a constant temperature of 37°C and 250rpm for 12-16h, and then place it at 4°C refrigerator cooling;

[0136] (2) After culturing the bacteria solution for 12-16 hours, measure its OD600 value with an ultraviolet spectrophotometer. If the OD600 value is between 0.6-0.8, it indicates that the bacteria are in the logarithmic growth phase of vigorous growth, and can be used for plasmid extraction. carry;

[0137] (3) Take the above cultured bacteria into a 50ml centrifuge tube, centrifuge at 5000rpm for 1 minute, collect the precipitate, and discard t...

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Abstract

The invention discloses a preparation method of a periodic Wuchereria malayi compound polyvalent protein vaccine. The preparation method comprises the steps of extraction of recombinant plasmids of the periodic Wuchereria malayi pcDNA3.1(+)-BmCPI / BmGAPDH, gene transfection, screening of positive clones, extraction and enlarge cultivation of the positive clones, cryopreservation of the positive clones, detection of the positive clones, extraction of cell total RNA (Ribose Nucleic Acid), molecular level detection of expression of the recombinant plasmids in cells, protein level detection of expression of the recombinant plasmids in cells, SDS-PAGE (Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis) analysis of expression products, extraction and purification of periodic Wuchereria malayi recombinant protein, and the like. The preparation method is simple and convenient, and good in effect.

Description

technical field [0001] The invention relates to a preparation method of a periodic type malayia filarial compound multivalent protein vaccine. Background technique [0002] The prevalence of filariasis not only brings serious harm to people's health, but also becomes one of the important causes of poverty due to disease. At present, the main method of treating filariasis is to use chemical drugs. Although drugs can reduce the infection rate and morbidity rate, due to frequent application, the risk of potential drug resistance of filariasis cannot be ruled out. In addition, repeated chemotherapy and long-term monitoring measures are required. Expensive fees and a large number of professional and technical personnel. Therefore, in order to find auxiliary or alternative methods to control filariasis, research on filariae vaccines has increasingly become an issue of concern. [0003] Although nucleic acid vaccines have shown great potential in the prevention and treatment of i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85C07K14/435A61K39/00C12N15/66A61P33/10
CPCY02A50/30
Inventor 方政王慧方浩陆施娟徐倩胡迎青童海燕徐邦生
Owner NANTONG UNIVERSITY
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