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Functional peptide of milk agglutinin and method for preparing same

A technology of lectin and functional peptide, which is applied in the field of biomedicine and infant health care, can solve the problem of difficult source of raw materials and achieve the effect of retaining antigenicity and high purity

Inactive Publication Date: 2007-08-15
XINHUA HOSPITAL ATTACHED TO SHANGHAI NO 2 MADICALUNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] The purpose of the present invention is to overcome the deficiencies of the prior art, and propose a lactagglutinin functional peptide and its preparation method, which (1) does not need to ensure fresh milk without any treatment, and only needs to preserve the recombinant DNA to be repeated Obtain fresh and active protein, which solves the problem of difficult source of raw materials; (2) Obtain a target protein with reproducible extraction, sufficient quantity, high purity, non-toxicity and antigenicity retention

Method used

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  • Functional peptide of milk agglutinin and method for preparing same
  • Functional peptide of milk agglutinin and method for preparing same
  • Functional peptide of milk agglutinin and method for preparing same

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Embodiment 1

[0037] 1. Materials:

[0038] 13 kinds of tissues were supplied by Shanghai Ruixing Gene Technology Co., Ltd.; PCR-Kit was purchased from TAKARA Company; Taq enzyme and DNA molecular weight standard were purchased from Shanghai Bioengineering Company; restriction endonuclease and T4 ligating enzyme were purchased from NEB Company;

[0039] Strains and plasmids: Transformed cells (DHa-MAX, BL-21) were purchased from Novagen; pET-28a was purchased from Promega;

[0040] PCR primers were synthesized by Shanghai Ruixing Gene Technology Co., Ltd.

[0041] Reagents: ITPG was purchased from Sigma Company; affinity chromatography column was purchased from Roche Company; His-tag antibody was purchased from Pharmingen Company.

[0042] 2. Method:

[0043] (1) Extraction of total RNA from 13 tissue mixtures

[0044] 1) Take 13 tissue mixtures (see Table 1), wash with TES, centrifuge and discard the supernatant;

[0045] 2) Add 2ml Trizol buffer and shake vigorously and transfer to a ...

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Abstract

The invention discloses a functional peptide of milk agglutinin and method for preparation, which comprises extracting total RNA from mixture 13 human tissues as the first chain of the cDNA, hooking total RNA from MGEF8A gene, and preparing the gene product of human milk agglutinator functional peptide by utilizing RT-PCR technology and cDNA PCR. The invention prepares destination proteins by means of protein engineering, the destination proteins obtained by the method have the advantages of repeated extraction, sufficient content, high purity, non-toxicity, and sustaining antigen property.

Description

technical field [0001] The invention belongs to the field of biomedicine, and can be applied to resisting rotavirus infection, regulating the immune function of the gastrointestinal tract of infants and the function maturation of intestinal epithelial cells, and the field of infant health care. Background technique [0002] A large number of epidemiological investigations have shown that rotavirus infection is the most important cause of diarrhea in children all over the world. Approximately 870,000 children under the age of 5 die from the disease each year in developing countries, accounting for 25% of deaths from diarrheal diseases in children under 5 years of age and 6% of all deaths from all causes. Among them, rotavirus Group A is the most widespread type in humans, and it is estimated that the number of deaths caused by Group A is about 800,000 per year. According to the survey, children under the age of 5 in my country suffer from 250 million diarrheal diseases every...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/09C12N15/63C07K14/435
Inventor 何振娟裴新红吴圣楣朱建幸杨凌云
Owner XINHUA HOSPITAL ATTACHED TO SHANGHAI NO 2 MADICALUNIV
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