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Grass carp FoxO1 polypeptide antigen and antibody and preparation method and application thereof

A technology of polypeptide antigen, foxo1, which is applied in the field of biochemistry and molecular immunology, can solve problems such as lack and achieve the effect of rapid preparation

Active Publication Date: 2019-04-19
XINXIANG MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The present invention also provides an antibody against grass carp FoxO1 polypeptide antigen, which can specifically recognize FoxO1 full-length functional protein in grass carp tissue, and can be used to detect the expression level of FoxO1 full-length functional protein in grass carp fat, liver and muscle tissue , solve the problem of lack of commercial grass carp FoxO1 antibody in the prior art, and fill the gap in the field of grass carp FoxO1 full-length functional protein detection

Method used

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  • Grass carp FoxO1 polypeptide antigen and antibody and preparation method and application thereof
  • Grass carp FoxO1 polypeptide antigen and antibody and preparation method and application thereof
  • Grass carp FoxO1 polypeptide antigen and antibody and preparation method and application thereof

Examples

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Embodiment 1

[0035] The grass carp FoxO1 polypeptide antigen in this example has the amino acid sequence shown in SEQ ID NO.1.

[0036] The process of obtaining the above-mentioned grass carp FoxO1 polypeptide antigen is as follows:

[0037] 1) Design and synthesis of grass carp FoxO1 polypeptide: the grass carp FoxO1 protein sequence was obtained according to the FoxO1 sequence on GenBank (accession number: ALI97589), which contains 654 amino acids (as shown in SEQ ID NO.3).

[0038] 2) Use DNAstar software to analyze the characteristics of grass carp FoxO1 protein. The analysis results are shown in Table 1. The protein has a molecular weight of 69444.26 Daltons and an isoelectric point of 6.408, which is an acidic protein.

[0039] Table 1 Characteristics of grass carp FoxO1 protein

[0040] Analysis project (Analysis)

Whole Protein

Molecular Weight

69444.26 Daltons

Length

654

Isoelectric Point

6.408

Charge (Charge at pH7)

-5.989 ...

Embodiment 2

[0044] The grass carp FoxO1 polypeptide antigen in this example is a C-terminally modified grass carp FoxO1 polypeptide antigen, which has the amino acid sequence shown in SEQ ID NO.2. The sequence is formed by linking a cysteine ​​to the C-terminus of the amino acid sequence shown in SEQ ID NO.1. The sequence of the polypeptide antigen is relatively short, so its immunogenicity is weak, and the addition of cysteine ​​is beneficial to the interaction between the polypeptide antigen and The carrier protein (maleamide-activated keyhole limpet hemocyanin KLH) is coupled to enhance the immunogenicity of the polypeptide antigen.

[0045] The above C-terminally modified grass carp FoxO1 polypeptide antigen was submitted to Shanghai Sangon Biotech Company for synthesis.

[0046] The identification of the grass carp FoxO1 polypeptide antigen in this embodiment includes the following steps:

[0047] 1) Apply HPLC, C184.6×250mm Alltima TM The separation column was analyzed, and gradie...

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Abstract

The invention relates to a grass carp FoxO1 polypeptide antigen and an antibody and a preparation method and application thereof, and belongs to the technical field of biochemistry and molecular immunology. According to the invention, the secondary structure, immunogenicity, hydrophilicity, hydrophobicity, surface probability and the like of a full-length functional protein amino acid sequence ofthe grass carp FoxO1 are analyzed, it is found that 15 amino acids of the grass carp FoxO1 polypeptide antigen, close to the N end, are strong in antigenicity, hydrophilicity and surface probability,therefore the 15 amino acids are used as the suitable grass carp FoxO1 polypeptide antigen. The polyclonal antibody for resisting the grass carp FoxO1 polypeptide antigen is prepared by immunizing NewZealand rabbits by taking the FoxO1 polypeptide antigen as an immunogen. The polyclonal antibody has the advantages of specifically recognizing the FoxO1 full-length functional protein in grass carptissues, filling in the blanks in the field of detection and research of the FoxO1 full-length functional protein of the grass carp, and laying a foundation for the research of regulating the fat metabolism function of an animal by the full-length functional protein of the grass carp FoxO1.

Description

technical field [0001] The invention relates to a grass carp FoxO1 polypeptide antigen, an antibody and a preparation method and application thereof, belonging to the technical fields of biochemistry and molecular immunology. Background technique [0002] Adipose tissue is the largest energy storage place in animals and plays a key role in energy balance in the body. Unbalanced development of adipose tissue often leads to many health problems. The location of fat deposition may be determined by tissue-specific lipid metabolism pathways, which are regulated by key enzymes and genes at the molecular level. Therefore, it is imperative to use molecular biology techniques to study the molecular mechanism of functional genes regulating fat metabolism. [0003] Grass carp is a unique freshwater fish in my country. It belongs to one of the "four major fishes" and has high economic value. Most farmed fish (especially grass carp) will experience problems such as fat accumulation, li...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/46C07K16/18C07K1/22G01N33/68
CPCC07K14/461C07K16/18G01N33/68
Inventor 张伟王明永周贝白姗姗刘友才史世强
Owner XINXIANG MEDICAL UNIV
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