Recombinant Wzt protein rabbit serum polyclonal antibody and preparation method thereof

A technology of polyclonal antibody and rabbit serum, which is applied to peptide preparation methods, botany equipment and methods, biochemical equipment and methods, etc., can solve problems such as LPS synthesis blockage

Inactive Publication Date: 2017-10-10
JILIN AGRICULTURAL UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Studies have found that the deletion of wzt gene can cause the synthesis of LPS to be blocked, bu...

Method used

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  • Recombinant Wzt protein rabbit serum polyclonal antibody and preparation method thereof
  • Recombinant Wzt protein rabbit serum polyclonal antibody and preparation method thereof
  • Recombinant Wzt protein rabbit serum polyclonal antibody and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0061] 1. Materials

[0062] 1. Strains and plasmids

[0063] The wzt gene was synthesized by Shanghai Shenggong Co., Ltd.

[0064] The pMD19T (Simple) vector was purchased from Bao Biological (Dalian) Company.

[0065] Template pET21b-wzt, prokaryotic expression vector pET-22b, DH5α clone competent cells and BL21(DE3) competent cells were preserved by the Laboratory of Metabolic Engineering and Synthetic Biology, College of Life Sciences, Jilin Agricultural University.

[0066] 2. Main reagents

[0067]

[0068] 3. Main instruments

[0069]

[0070]

[0071] 2. Preparation of recombinant Wzt protein

[0072] 1. Construction of recombinant cloning plasmid pMD-19T-wzt

[0073] (1) Preparation of Escherichia coli DH5α competent cells

[0074] ① Take out the original strain of E.coli DH5α from the -80°C refrigerator, streak culture on the LB solid plate, pick a single colony, inoculate it in 5mL LB liquid medium, and cultivate it at 37°C, 150rpm, shaking for 12h, a...

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Abstract

The invention relates to a recombinant Wzt protein rabbit serum polyclonal antibody and a preparation method thereof, which relate to the field of antibody preparation, and fill the gap of applying recombinant Wzt protein for preparing the specific polyclonal antibody. The preparation method of the polyclonal antibody comprises the steps of after mixing and emulsifying the recombinant Wzt protein and an equivoluminal freund's complete adjuvant, carrying out multiple sites subcutaneous injection on the back parts of female rabbits; carrying out secondary immunizing: after one week, after mixing and emulsifying the recombinant Wzt protein and the equivoluminal freund's incomplete adjuvant, carrying out multiple sites subcutaneous injection on the back parts of the female rabbits; carrying out thrice immunizing and quartic immunizing every other week according to a secondary immunizing process; collecting venous blood of female domestic rabbit ears after immunizing each time, and detecting a rabbit serum antibody titer; at one week after quartic immunizing, collecting rabbit whole blood, centrifugally collecting a supernatant to obtain a rabbit serum, and obtaining the recombinant Wzt protein rabbit serum polyclonal antibody after purifying. The recombinant Wzt protein rabbit serum polyclonal antibody has favorable specificity, and can be used for detecting Brucella Wzt protein.

Description

technical field [0001] The invention relates to the technical field of antibody preparation, in particular to a recombinant Wzt protein rabbit serum polyclonal antibody and a preparation method thereof. Background technique [0002] Brucellosis (brucellosis, referred to as brucellosis) is caused by Brucella, mainly infecting ruminants and humans, and can also infect deer, dogs, etc. Brucellosis is infected in most parts of the world reports. [0003] Animals suffering from brucellosis can cause a series of problems such as abortion, reduced milk production and infertility in pregnant animals, which has a very large economic impact, especially in developing countries. The vast majority of Brucella can infect people and cause serious diseases, not only need to cooperate with the treatment of antibiotics, but also the treatment time is relatively long, and the sequelae left behind are usually relatively serious. The spread of brucellosis has seriously affected the development...

Claims

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Application Information

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IPC IPC(8): C07K16/12C07K16/06C12N15/70C12N15/31C12N15/66C07K14/23C07K1/22
CPCC07K14/23C07K16/06C07K16/1221C12N15/66C12N15/70C12N2800/22
Inventor 王秀然王浩卢天成王岩李海燕付玉芹孙喆李思圆贾宇王茗宇刘思阳胡祥坤边红李全亮
Owner JILIN AGRICULTURAL UNIV
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