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Preparation of serologic H-Y antigen gene DBY antibody and XY sperm immune sorting method thereof

An antigen gene, antigen immunization technology, applied in the field of genetic engineering, can solve the problem of difficulty in preparing DBY-specific antibodies

Active Publication Date: 2014-06-11
HUNAN AGRICULTURAL UNIV
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  • Description
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AI Technical Summary

Problems solved by technology

[0006] Due to the high homology between the DBY gene and the corresponding gene Dbx on the X chromosome, it is very difficult to prepare DBY-specific antibodies

Method used

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  • Preparation of serologic H-Y antigen gene DBY antibody and XY sperm immune sorting method thereof
  • Preparation of serologic H-Y antigen gene DBY antibody and XY sperm immune sorting method thereof
  • Preparation of serologic H-Y antigen gene DBY antibody and XY sperm immune sorting method thereof

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Embodiment Construction

[0041] The present invention is further described below in conjunction with specific embodiments, rather than limiting the present invention.

[0042] 1. Bioinformatics prediction and screening of DBY antigen peptides

[0043] Use DNAStar software PROTEAN program (Gamier-Robson program), online protein analysis tool ABCpred servers ( http: / / www.imtech.res.in / raghava / abcpred / ), BepiPred1.0servers (http: / / www.cbs.dtu.dk / services / BepiPred) and other bioinformatics technologies, respectively from the protein transmembrane region, primary structure homology, secondary structure, hydrophilicity, In terms of plasticity, surface polarity, and antigenic index, we comprehensively analyze the structural characteristics of the entire DBY protein sequence, and we also predict the transmembrane region of the DBY protein. The DBY of the male individual is aligned with the DBX of the female individual to remove redundant sequences and homologous repetitive sequences to further narrow the predic...

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Abstract

The invention discloses the preparation of a serologic H-Y antigen gene DBY antibody and an XY sperm immune sorting method thereof. The method comprises the following steps of: predicting and screening three B cell epitope antigen peptides of the H-Y antigen gene DBY antibody by a method in information biology, and connecting, recombining and performing prokaryotic expression to obtain high-purity DBY recombinant antigen peptide; immunizing a New Zealand white rabbit based on the recombinant antigen peptide as an antigen to obtain a polyclonal antibody high in valence and strong in specificity; and sorting about 63.2% of sperm, 4.5% plus or minus by DBY antibody fixation flow cytometry. According to the invention, the XY immune sorting method is established based on the serologic H-Y antigen gene DBY antibody, so that a simple, efficient and safe ideal sex selection method can be established by separating X and Y sperms by the antibody, therefore, genetic improvement of seed variety (or variety) is better controlled so as to obtain faster genetic progress and efficiency advantages on production and management in the animal husbandry.

Description

Technical field [0001] The invention belongs to the technical field of genetic engineering, and specifically relates to a serological H-Y antigen gene DBY recombinant antigen peptide, and a serological H-Y antigen gene DBY antibody preparation and XY sperm immune sorting method. Background technique [0002] Male specific minor histompatibility antigens (H-Y antigens) were originally discovered as a heterosexual transplantation antigen, a substance specific to male animals, and no tissue and organ specificity in male individuals. With the deepening of research, it was discovered that H-Y antigen is a glycoprotein that exists on the surface of male mammalian cells. HY antibody is of great significance in the theory and practice of mammalian sex identification and control. Many laboratories at home and abroad have produced HY antigen antiserum and monoclonal antibodies, and performed embryo sex identification and control, but due to its antigen The specificity is weak and complex,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/14C07K16/40G01N33/569C12N5/076
Inventor 王乃东谭庆辉薛立群袁安文许道军
Owner HUNAN AGRICULTURAL UNIV
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