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Function and use of MNSF-beta gene during process of embryo nidation

An embryo and implantation technology, applied in the fields of biotechnology and medicine, can solve the problems of poorly understood effects and not directly exerting immunosuppressive effects

Inactive Publication Date: 2010-05-12
SHANGHAI INST OF PLANNED PARENTHOOD RES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] To sum up, at present, people know little about the influence of MNSFβ gene and its expression products on embryo implantation and pregnancy, and it is believed that it does not directly play an immunosuppressive role in the process of embryo implantation
However, the above research results are only superficial speculations, and it is necessary to further study the mechanism of action of the MNSFβ gene during embryo implantation and pregnancy

Method used

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  • Function and use of MNSF-beta gene during process of embryo nidation
  • Function and use of MNSF-beta gene during process of embryo nidation
  • Function and use of MNSF-beta gene during process of embryo nidation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Expression of MNSFβ protein in mouse oocytes and early embryos before implantation

[0065] The inventors performed immunofluorescence analysis on oocytes, 2-cell stage embryos and blastocysts. see results figure 1 , it was found that positive signals of MNSFβ protein were detected in egg cells (1), 2-cell stage embryos (2) and blastocysts (3).

[0066] Since the exposure coefficient is fixed, the strength of the signal can reflect the expression level of the protein to be tested to a certain extent. Therefore, according to the fluorescence intensity, it is speculated that the expression level of MNSFβ protein in early embryos may increase with pregnancy.

Embodiment 2

[0068] Antibody blocking experiment confirms that anti-MNSFβ antibody can specifically inhibit mouse embryo implantation

[0069] In this example, the complete ORF coding sequence of MNSFβ was obtained by conventional PCR method, and then inserted into the multiple cloning site of pGEX-4T-2 (purchased from Amersham Pharmacia Biotech Company), thereby obtaining the expression plasmid pGEX-4T-2 / MNSFβ. Use this plasmid to express GST-MNSFβ fusion protein in Escherichia coli; then treat the purified GST-MNSFβ fusion protein with bovine thrombin, isolate the recombinant MNSFβ protein, and use it as an immunogen, using Freund’s adjuvant as an immunostimulator , immunized rabbits according to conventional methods, and prepared high-titer rabbit anti-MNSFβ serum; and then separated biologically active recombinant MNSFβ (rMNSFβ) from the antiserum through Protein A Sepharose CL-4B column chromatography, and effectively Rabbit anti-MNSFβ-IgG that neutralizes the biological activity of...

Embodiment 3

[0089] Example 3 Analysis of the Molecular Mechanism of Anti-MNSFβ Antibody Inhibiting Embryo Implantation Using cDNA Chip Technology

[0090]In order to further understand the biological function of the MNSFβ gene during pregnancy, the inventors applied the cDNA chip technology and used the Affymetrix MOUSE 430A gene expression profile chip to analyze the pair of anti-MNSFβ antibody-treated and non-anti-MNSFβ antibody-treated Between the uterine tissues, the difference in gene expression was analyzed, and the results were scanned with the Affymetrix gene chip scanner, and the data were read and processed with the microarray analysis software Microarray Suite5.0 (Affymetrix company), and the differential gene screening standard was : "Change" is I or MI, Ratio> or = 1, "experimental group" Detection is "P" as up-regulated genes; "Change" is D or MD, Ratio figure 2 , image 3 ).

[0091] The present inventors have screened out 144 known genes with expression differences betwee...

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Abstract

An application of MNSF beta gene or its coding protein in regulatin the nidation of embryo, and the application of the antagon of MNSF beta (such as the specific antibody or antisense nucleic acid ofMNSF) in specifically suppressing the nidation of embryo are disclosed.

Description

technical field [0001] The invention relates to the fields of biotechnology and medicine, in particular, the invention relates to the function and application of the MNSFβ gene in the process of embryo implantation. Background technique [0002] MNSF (monoclonal nonspecific suppressor factor, monoclonal nonspecific suppressor factor) was originally found to be a lymphocyte factor with nonspecific immunosuppressive effect produced by mouse T cell hybridoma (Nakamura M et al., (1986), Isolation and Characterization of amonoclonal nonspecific suppressor factor (MNSF) produced by a T cell hybridoma. Journal of Immunology, 136: 2904-2909). And MNSFβ is a subunit of MNSF, and it is also the most important component of MNSF to exert its biological function (Nakamura M et al., (1996), Ubiquitin-like moiety of the monoclonal nonspecific suppressor factor beta is responsible for its activity. Journal of Immunology, 156: 532-538), can inhibit the synthesis and secretion of IgG, and th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61K38/19A61P15/18G01N33/68C12Q1/68
Inventor 王健申庆祥黄哲平
Owner SHANGHAI INST OF PLANNED PARENTHOOD RES
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