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Functional peptide of milk agglutinin separated from MCF-7 and method for preparing same

A lactagglutinin and MCF-7 technology, applied in the fields of infant health care and biomedicine, can solve problems such as difficulty in the source of raw materials, and achieve the effect of retaining antigenicity and high purity

Inactive Publication Date: 2007-10-10
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 extracted from MCF-7 and a preparation method. It (1) does not need to ensure fresh milk without any treatment, and only needs to preserve the recombinant The fresh and active protein can be repeatedly obtained from the DNA, 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 separated from MCF-7 and method for preparing same
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  • Functional peptide of milk agglutinin separated from MCF-7 and method for preparing same

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Embodiment 1

[0036] 1. Materials:

[0037] MCF-7 breast cancer cells were purchased from the Institute of Cells, Chinese Academy of Sciences; PCR-Kit was purchased from TAKARA Company; Taq enzymes and DNA molecular weight standards were purchased from Shanghai Bioengineering Company; restriction enzymes and T4 ligating enzymes were purchased from NEB Company;

[0038] Strains and plasmids: Transformed cells (DHa-MAX, BL-21) were purchased from Novagen; pET-28a was purchased from Promega;

[0039] PCR primers were synthesized by Shanghai Ruixing Gene Technology Co., Ltd.

[0040] Reagents: ITPG was purchased from Sigma Company; affinity chromatography column was purchased from Roche Company; His-tag antibody was purchased from Pharmingen Company.

[0041] 2. Method:

[0042] (1) Extraction of total RNA from cells

[0043] 1) Take 1×10 MCF-7 breast cancer cells 7 Cells were washed with TES, and the supernatant was discarded by centrifugation;

[0044] 2) Add 1ml Trizol buffer solution 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 MCF-7 mammary cancer cell strains, synthesizing template cDNA through RT-PCR technology, and preparing gene product of human milk agglutinin functional peptide. 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 properety.

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): C07K14/435C07K14/47C12N15/09C12N15/63A61P31/12
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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